affy/DESCRIPTION0000644000175200017520000000463014516022632014233 0ustar00biocbuildbiocbuildPackage: affy Version: 1.80.0 Title: Methods for Affymetrix Oligonucleotide Arrays Author: Rafael A. Irizarry , Laurent Gautier , Benjamin Milo Bolstad , and Crispin Miller with contributions from Magnus Astrand , Leslie M. Cope , Robert Gentleman, Jeff Gentry, Conrad Halling , Wolfgang Huber, James MacDonald , Benjamin I. P. Rubinstein, Christopher Workman , John Zhang Maintainer: Robert D. Shear URL: https://bioconductor.org/packages/affy BugReports: https://github.com/rafalab/affy/issues Depends: R (>= 2.8.0), BiocGenerics (>= 0.1.12), Biobase (>= 2.5.5) Imports: affyio (>= 1.13.3), BiocManager, graphics, grDevices, methods, preprocessCore, stats, utils, zlibbioc Suggests: tkWidgets (>= 1.19.0), affydata, widgetTools, hgu95av2cdf LinkingTo: preprocessCore Description: The package contains functions for exploratory oligonucleotide array analysis. The dependence on tkWidgets only concerns few convenience functions. 'affy' is fully functional without it. License: LGPL (>= 2.0) Collate: ProgressBarText.R ppset.ttest.R ppsetApply.R expressoWidget.R getCDFenv.R AffyRNAdeg.R avdiff.R barplot.ProbeSet.R bg.Affy.chipwide.R bg.R expresso.R fit.li.wong.R generateExprVal.method.avgdiff.R generateExprVal.method.liwong.R generateExprVal.method.mas.R generateExprVal.method.medianpolish.R generateExprVal.method.playerout.R hlog.R justrma.R loess.normalize.R maffy.R mas5.R merge.AffyBatch.R normalize.constant.R normalize.contrasts.R normalize.invariantset.R normalize.loess.R normalize.qspline.R normalize.quantiles.R pairs.AffyBatch.R plot.density.R plotLocation.R plot.ProbeSet.R pmcorrect.mas.R AffyBatch.R mva.pairs.R ProbeSet.R read.affybatch.R rma.R summary.R tukey.biweight.R whatcdf.R xy2indices.R zzz.R biocViews: Microarray, OneChannel, Preprocessing LazyLoad: yes git_url: https://git.bioconductor.org/packages/affy git_branch: RELEASE_3_18 git_last_commit: a0d64df git_last_commit_date: 2023-10-24 Date/Publication: 2023-10-24 NeedsCompilation: yes Packaged: 2023-10-24 20:14:18 UTC; biocbuild affy/NAMESPACE0000644000175200017520000000452514516003533013746 0ustar00biocbuildbiocbuilduseDynLib("affy") importFrom(BiocGenerics, updateObject, colnames, boxplot, image) import(affyio) import(zlibbioc) importClassesFrom(Biobase, AnnotatedDataFrame, AssayData, eSet, MIAME, Versioned, VersionedBiobase, Versions) importClassesFrom(methods, ANY, character, environment, integer, matrix, missing, numeric) importMethodsFrom(Biobase, annotatedDataFrameFrom, annotation, assayData, classVersion, "classVersion<-", description, "description<-", experimentData, exprs, "exprs<-", featureNames, "featureNames<-", isCurrent, isVersioned, notes, "notes<-", pData, "pData<-", phenoData, "phenoData<-", preproc, "preproc<-", protocolData, "protocolData<-", rowMedians, sampleNames, "sampleNames<-", se.exprs, "se.exprs<-") importMethodsFrom(methods, initialize, show) importFrom(Biobase, addVigs2WinMenu, assayDataElementReplace, assayDataNew, multiassign, read.AnnotatedDataFrame, read.MIAME, testBioCConnection) importMethodsFrom(BiocGenerics, normalize) importFrom(BiocManager, repositories) importFrom(graphics, abline, axis, barplot, frame, hist, lines, matplot, mtext, pairs, par, plot, points, smoothScatter, text) importFrom(grDevices, dev.interactive, gray) importFrom(methods, as, callNextMethod, getMethod, is, new, slot) importFrom(preprocessCore, normalize.quantiles, normalize.quantiles.robust, rma.background.correct) importFrom(stats, approx, density, IQR, lm, loess, loess.control, median, medpolish, model.response, model.weights, optim, pnorm, predict, quantile, runif, sd, smooth.spline, splinefun, t.test, terms, var, wilcox.test) importFrom(utils, available.packages, contrib.url, data, flush.console, install.packages, object.size, packageVersion) ##export everything that does not start with a . exportPattern("^[^\\.]") export(.setAffyOptions) exportMethods(show, boxplot, image, exprs, "exprs<-", featureNames, geneNames, sampleNames, se.exprs, updateObject) exportClasses("AffyBatch") ## S3 methods S3method(plot, ProbeSet) S3method("$", AffyBatch) S3method(barplot, ProbeSet) S3method(merge, AffyBatch) S3method(pairs, AffyBatch) affy/NEWS0000644000175200017520000001247214516003533013226 0ustar00biocbuildbiocbuildversion 1.41.1 o Fixed bug. attr(r,"constant") <- thisconstant * refconstant to attr(r,"constant") <- refconstant / thisconstant. Reported Aris Floratos. version 1.23.x o Populate new eSet slot 'protocolData' with information from cel files. version 1.17.x o Replaced usage of defunct Biobase classes exprSet and phenoData with ExpressionSet and AnnotatedDataFrame. version 1.4.x [under development] o xy2indices/indices2xy have now a (dangerous) parameter xy.offset. See below for more details. o a new package-wide parameter called 'xy.offset' was added. This is the first towards getting rid of the 'indexing-starting-at-one-and -not-at-zero' difficulty. This package-wide parameter is the only thing one should play play with to change the offset of x/y coordinates for features on a chip. o computeExprSet (method of AffyBatch) has the following (visible) improvements: - better reporting of errors - better handlings ids as parameters (does not crash any longer when unknown ids are given) o indexProbes (method of AffyBatch) sees the deprecated flag 'xy' removed for good. o mmindex (see indexProbes above) o pmindex (see indexProbes above) o ppsetApply: a function to apply a function over the probe sets that can be built from an instance of AffyBatch. This is done in the spirit of esetApply in Biobase (the covariate information in phenoData are directly accessible from the function (see example in the man page). o rma - the subset parameter now works properly. o mva.pairs - now uses a subset to fit the loess curve (much more efficient). also the summary statistics displayed are different. In particular the median and IQR of the M's are shown in the lower triangle. Previously, an IQR of loess curve values was shown. version 1.3.x Things done: 0 (wh:) moved everything related to Cdf-class from 'affy' to 'makecdfenv': R/Cdf.R, R/getLocationsData.Cdf.R, locate.name.R, pmormm.R R/read.cdffile.R, data/cdf.example.rda, man/Cdf-class.Rd man/getLocationsData.Cdf.Rd, man/locate.name.Rd, man/pmormm.Rd man/read.cdffile.Rd, src/read_cdffile.c o read.affybatch2 reads all cel files together in one big hit into an affybatch o the baseline in normalize.AffyBatch,invariant can be selected using a baseline.type parameter. Also changed "progress" to the more standard "verbose". Seemed to be an error since both PM and MM probes were normalized by only PM's where used to establish the normalization, now follows same standard as been introduced for normalize.AffyBatch.quantiles. o Most normalization routines can now be called with the parameter type which specifies whether to be pmonly, mmonly, both together or both separately. This introduces more consistency to the how the normalization routines are actually applied to affybatchs. o express() has been completely removed. Things that would be good to see before next release: o More consistent usage/application of MIAME version 1.2.x: o Autoload of cdfenvs on demand (uses reposTools). Can be configured through the options. o slot 'preprocessing' of the MIAME attribute used to store normalization step information [will be polished, list returned for the moment. Need for a class and check compliance with MIAME standards] o default methods for normalization, bg correction, pm correction and summary now in the package options [options exist for all, but only used by normalize for the moment]. o tuning of the MAS5.0 methods implemented (bgcorrect.mas, ...) [Ben for details. add URL for the comparison he made] o method plot.ProbeSet, an alternative to barplot, to plot probe level information. o parameter 'scale' in the method barplot for ProbeSet. All the barplots are scaled to eachothers.This is the default. o bug in the parser fixed (infinite loop reported with apparently non-standard CEL files.). o bug in the parser fixed (the 'sd' data returned were not correct). o missing slot in the dataset SpikeIn fixed. o The function express() is deprecated. It still functions normally but gives warning mesage. It will be removed in a future release. The function expresso() should be used as a replacement. o bug in normalize.AffyBatch.qspline fixed (thanks to people at Insightful). The expression data matrix sent to normalize.qspline was mistakingly transposed. o The default background on the rma() function has been changed. Now the results from rma() and expresso() should agree completely. o New functions 'xy2indices' and 'indices2xy' to shuttle from x/y pos to indices (like the ones in cdfenvs) (and reverse). o Reformating of the documentation. version 1.1.x: o 'image(cel)' scales to the size (# rows and # cols) of a chip. locations can be plotted over directly ('plotLocations' was fixed accordiginly). o 'write.celfile(cel)' to write Cel objects into .CEL files o 'getInfoInAffyFile' to snoop in CEL and CDF files o 'whatcdf' to get the name of the CDF from the CEL o one more slot in class 'Cdf': cdfName (will match with what is returned by 'whatcdf') o one more slot in class 'Cel': cdfName (returned by whatcdf) o new class 'AffyBatch': cdfName (returned by whatcdf) o extensive set of tests in the directory 'tests/' affy/R/0000755000175200017520000000000014516003533012722 5ustar00biocbuildbiocbuildaffy/R/AffyBatch.R0000644000175200017520000007261614516003533014710 0ustar00biocbuildbiocbuild##RG added a method to store and update the available normalization methods normalize.AffyBatch.methods <- function() .affyInternalEnv[["normalize.AffyBatch.methods"]] upDate.normalize.AffyBatch.methods <- function(x) { if (is.character(x)) .affyInternalEnv[["normalize.AffyBatch.methods"]] <- x } express.summary.stat.methods <- function() .affyInternalEnv[["express.summary.stat.methods"]] upDate.express.summary.stat.methods <- function(x) { if (is.character(x)) .affyInternalEnv[["express.summary.stat.methods"]] <- x } generateExprSet.methods <- function() .affyInternalEnv[["generateExprSet.methods"]] upDate.generateExprSet.methods <- function(x) { if (is.character(x)) .affyInternalEnv[["generateExprSet.methods"]] <- x } bgcorrect.methods <- function() .affyInternalEnv[["bgcorrect.methods"]] upDate.bgcorrect.methods <- function(x) { if (is.character(x)) .affyInternalEnv[["bgcorrect.methods"]] <- x } pmcorrect.methods <- function() .affyInternalEnv[["pmcorrect.methods"]] upDate.pmcorrect.methods <- function(x) { if (is.character(x)) .affyInternalEnv[["pmcorrect.methods"]] <- x } ## computeExprSet: ## - better reporting of errors ## - better handling of ids (does not crash any longer when unknown id) ## - use of the progress bar in Biobase 1.4.4 ## - cleanup of the comments in the code ## indexProbes: ## - deprecated flag 'xy' removed for good if (debug.affy123) cat("-->initAffyBatch\n") ## Inherits from Affybatch ## The accessor 'intensity' gets what is in the slot 'exprs' setClass("AffyBatch", representation=representation( cdfName="character", nrow="numeric", ncol="numeric"), contains="eSet", prototype=prototype( new("VersionedBiobase", versions=c(classVersion("eSet"), AffyBatch="1.2.0")))) setMethod("initialize", signature(.Object="AffyBatch"), ## provide a signature consistent with the new defintion, but that picks up the (implicit) old function(.Object, cdfName="", nrow=0, ncol=0, phenoData, featureData, experimentData=new("MIAME"), annotation=character(0), protocolData=phenoData[,integer(0)], assayData, exprs=matrix(numeric(0), nrow=nrow, ncol=ncol), ## se.exprs ...) { .Object@cdfName <- cdfName .Object@nrow <- nrow .Object@ncol <- ncol dots <- list(...) ## remove reporterInfo, description, notes from '...' if ("reporterInfo" %in% names(dots)) { if (missing(featureData)) { if (!is.null(dots[["reporterInfo"]])) { if (is(dots[["reporterInfo"]], "data.frame")) { featureData <- new("AnnotatedDataFrame", data=dots[["reporterInfo"]]) } else { warning("trying to convert reporterInfo (class '", class(dots[["reporterInfo"]]), "') to featureData (class 'AnnotatedDataFrame')", immediate.=TRUE) featureData <- as(dots[["reporterInfo"]], "AnnotatedDataFrame") } } else { stop("use 'featureData' rather than 'reporterInfo' for feature covariates") } } } if ("description" %in% names(dots)) { if (missing(experimentData)) experimentData <- dots[["description"]] else stop("use 'experimentData' rather than 'description' for experiment description") } if ("notes" %in% names(dots)) { ## warning("addding 'notes' to 'experimentData'") notes(experimentData) <- c(notes(experimentData), dots[["notes"]]) } dots <- dots[!names(dots) %in% c("reporterInfo", "description", "notes")] ## update phenoData to AnnotatedDataFrame, if necessary if (missing(assayData)) { assayData <- do.call(assayDataNew, c(list(exprs=exprs), dots)) } else if (!missing(exprs)) { stop("cannot initialize when both 'assayData' and 'exprs' are specified") } if (missing(phenoData) || is.null(phenoData)) phenoData <- annotatedDataFrameFrom(assayData, byrow=FALSE) else if (!is(phenoData, "AnnotatedDataFrame")) phenoData <- as(phenoData, "AnnotatedDataFrame") if (missing(featureData)) featureData <- annotatedDataFrameFrom(assayData, byrow=TRUE) callNextMethod(.Object, assayData=assayData, phenoData=phenoData, featureData=featureData, experimentData=experimentData, annotation=annotation, protocolData=protocolData) }) setMethod("updateObject", signature(object="AffyBatch"), function(object, ..., verbose=FALSE) { if (verbose) message("updateObject(object = 'AffyBatch'") if (isVersioned(object) && isCurrent(object)["AffyBatch"]) return(callNextMethod()) if (!isVersioned(object) || is.null(classVersion(object)[["AffyBatch"]])) { exprs <- slot(object, "exprs") se.exprs <- slot(object, "se.exprs") if (!all(dim(se.exprs) == dim(exprs))) { if (!all(dim(se.exprs)==0)) warning("removing 'se.exprs' with dimensions different from 'exprs'") se.exprs <- NULL } if ("reporterInfo" %in% names(attributes(object)) && any(dim(slot(object, "reporterInfo"))!=0)) warning("reporterInfo data not transfered to 'AffyBatch' object") experimentData=updateObject(slot(object, "description")) if ("notes" %in% names(attributes(object)) && length(slot(object, "notes"))!=0) { warning("adding 'notes' to 'experimentData'") notes(experimentData) <- c(notes(experimentData), object@notes) } if (!is.null(se.exprs)) new("AffyBatch", phenoData=as(slot(object, "phenoData"), "AnnotatedDataFrame"), experimentData=experimentData, annotation=slot(object, "annotation"), cdfName=slot(object, "cdfName"), nrow=slot(object, "nrow"), ncol=slot(object, "ncol"), exprs=exprs, se.exprs=se.exprs) else new("AffyBatch", phenoData=as(slot(object, "phenoData"), "AnnotatedDataFrame"), experimentData=experimentData, annotation=slot(object, "annotation"), cdfName=slot(object, "cdfName"), nrow=slot(object, "nrow"), ncol=slot(object, "ncol"), exprs=exprs) } else if (!isCurrent(object)[["AffyBatch"]]) { ## remove empty se.exprs, introduced in class version 1.2.0 se.exprs <- assayData(object)[["se.exprs"]] if (!is.null(se.exprs) && all(is.na(se.exprs))) { message("removing 'se.exprs' with all NA values") se.exprs(object) <- NULL } object <- callNextMethod() classVersion(object)["AffyBatch"] <- classVersion("AffyBatch")["AffyBatch"] object } else object }) ####################################################### ### accessors ####################################################### if (debug.affy123) cat("--->accessors\n") setMethod("exprs", signature(object="AffyBatch"), function(object) assayData(object)[["exprs"]]) setReplaceMethod("exprs", signature(object="AffyBatch"), function(object, value) { assayDataElementReplace(object, "exprs", value, validate=FALSE) }) setMethod("se.exprs", signature(object="AffyBatch"), function(object) { obj <- assayData(object)[["se.exprs"]] if (is.null(obj)) new("matrix") else obj }) setReplaceMethod("se.exprs", signature(object="AffyBatch"), function(object, value) { assayDataElementReplace(object, "se.exprs", value, validate=FALSE) }) if (is.null(getGeneric("cdfName"))) setGeneric("cdfName", function(object) standardGeneric("cdfName")) setMethod("cdfName", "AffyBatch", function(object) object@cdfName) ##intensity setGeneric("intensity", function(object) standardGeneric("intensity")) setMethod("intensity", signature(object="AffyBatch"), function(object) assayData(object)[["exprs"]]) setGeneric("intensity<-", function(object, value) standardGeneric("intensity<-")) setReplaceMethod("intensity", signature(object="AffyBatch"), function(object, value) { assayDataElementReplace(object, "exprs", value, validate=FALSE) }) ##for now, there is no accessor for se.exprs. we could use this to store ##sd, but if no one uses it... why do it setMethod("length",signature(x="AffyBatch"), function(x) ncol(exprs(x))) ##RI: assumes matrices setMethod("dim", signature=signature(x="AffyBatch"), function(x) c(x@nrow, x@ncol)) ####################################################### ### methods ####################################################### setMethod("featureNames", signature=signature(object="AffyBatch"), function(object) { cdf.envir <- getCdfInfo(object) ls(envir=cdf.envir) }) setReplaceMethod("featureNames", signature=signature( object="AffyBatch", value="ANY"), function(object, value) stop("Cannot change featureNames of AffyBatch")) ##geneNames method if (debug.affy123) cat("--->geneNames\n") if( is.null(getGeneric("geneNames") )) setGeneric("geneNames", function(object) standardGeneric("geneNames")) setMethod("geneNames",signature("AffyBatch"), function(object){ cdf.envir <- getCdfInfo(object) return(ls(envir=cdf.envir)) }) ##show method if (debug.affy123) cat("--->show\n") setMethod("show", "AffyBatch", function(object) { if (!isVersioned(object) || classVersion(object)["AffyBatch"] < "1.1.0") stop("AffyBatch out-of-date; use 'updateObject()'", call.=FALSE) if (!isCurrent(object)['AffyBatch']) message("AffyBatch out-of-date; consider 'updateObject()'") ## Location from cdf env cdf.env <- tryCatch(getCdfInfo(object), error=function(err) { warning("missing cdf environment! in show(AffyBatch)", call.=FALSE) NULL }) num.ids <- if (!is.null(cdf.env)) length(ls(envir=cdf.env)) else num.ids <- "???" cat("AffyBatch object\n") cat("size of arrays=", nrow(object), "x", ncol(object), " features (", object.size(object) %/% 1024, " kb)\n", sep="") cat("cdf=", object@cdfName, " (", num.ids, " affyids)\n", sep="") cat("number of samples=", length(object), "\n", sep="") cat("number of genes=", length(featureNames(object)), "\n", sep="") cat("annotation=", object@annotation, "\n", sep="") if(length(notes(object)) > 0) cat("notes=",paste(notes(object),collapse="\n\t"), "\n", sep="") }) # if (is.null(getGeneric("index2xy"))) { # setGeneric("indexProbes", function(object, which, ...) # standardGeneric("indexProbes")) # } ## indexProbes if( is.null(getGeneric("indexProbes"))) setGeneric("indexProbes", function(object, which, ...) standardGeneric("indexProbes")) setMethod("indexProbes", signature=c(object="AffyBatch", which="missing"), function(object, which, ...) indexProbes(object, which="pm", ...)) setMethod("indexProbes", signature("AffyBatch", which="character"), function(object, which=c("pm", "mm","both"), genenames=NULL) { which <- match.arg(which) i.probes <- match(which, c("pm", "mm", "both")) ## i.probes will know if "[,1]" or "[,2]" ## if both then [,c(1,2)] if(i.probes==3) i.probes=c(1,2) envir <- getCdfInfo(object) if(is.null(genenames)) genenames <- ls(envir ) ## note: the variable name genenames could be confusing (the same gene can be ## found in several affyid (ex: the 3' and 5' controls) ans <- mget(genenames, envir, ifnotfound=NA) ## this kind of thing could be included in 'mget' as ## an extra feature. A function could be specified to ## process what is 'multi'-get on the fly for (i in seq(along=ans)) { if ( is.na(ans[[i]][1]) ) next ##as.vector cause it might be a matrix if both tmp <- as.vector(ans[[i]][, i.probes]) ans[[i]] <- tmp } return(ans) }) ##pmindex method if( is.null(getGeneric("pmindex"))) setGeneric("pmindex", function(object,...) standardGeneric("pmindex")) ##wrapper setMethod("pmindex", "AffyBatch", function(object, genenames=NULL) indexProbes(object, "pm", genenames=genenames)) ##mmindex method if( is.null(getGeneric("mmindex"))) setGeneric("mmindex", function(object,...) standardGeneric("mmindex")) ##wrapper setMethod("mmindex", "AffyBatch", function(object,genenames=NULL) indexProbes(object, "mm", genenames=genenames)) ##probeNames method if( is.null(getGeneric("probeNames"))) setGeneric("probeNames", function(object, ...) standardGeneric("probeNames")) setMethod("probeNames","AffyBatch", function(object, genenames=NULL, mm=FALSE){ if(mm) Index <- mmindex(object,genenames) else Index <- pmindex(object,genenames) reps <- unlist(lapply(Index,length),use.names=FALSE) rep(names(Index),reps) }) if( is.null(getGeneric("probes")) ) setGeneric("probes", function(object, ...) standardGeneric("probes")) setMethod("probes", signature("AffyBatch"), function(object, which=c("pm", "mm"), genenames=NULL, LISTRUE=FALSE, drop=FALSE){ which <- match.arg(which) index <- indexProbes(object, which, genenames) if(LISTRUE) ans <- lapply(index, function(i) exprs(object)[i, ,drop=drop]) else{ index <- unlist(index) ans <- exprs(object)[index, ,drop=drop] colnames(ans) <- sampleNames(object) rownames(ans) <- names(index) } return(ans) }) ##pm method if( is.null(getGeneric("pm") )) setGeneric("pm", function(object, ...) standardGeneric("pm")) setMethod("pm","AffyBatch", function(object, genenames=NULL, LISTRUE=FALSE){ if(is.null(genenames) & !LISTRUE){ cdfname <- getCdfInfo(object) psets<- as.list(cdfname) psets<- psets[order(names(psets))] index <-unlist(sapply(psets, function(x) x[,1]),use.names=FALSE) return(exprs(object)[index,,drop=FALSE]) } else{ return(probes(object, "pm", genenames, LISTRUE=LISTRUE)) } }) if( is.null(getGeneric("pm<-") )) setGeneric("pm<-", function(object, value) standardGeneric("pm<-")) setReplaceMethod("pm", "AffyBatch", function(object, value){ Dimnames <- dimnames(exprs(object)) cdfname <- getCdfInfo(object) psets<- as.list(cdfname) psets<- psets[order(names(psets))] pmIndex <-unlist(sapply(psets, function(x) x[,1]),use.names=FALSE) exprs(object)[pmIndex,] <- value dimnames(exprs(object)) <- Dimnames object }) ##mm method if( is.null(getGeneric("mm") )) setGeneric("mm", function(object, ...) standardGeneric("mm")) setMethod("mm",signature("AffyBatch"), function(object, genenames=NULL, LISTRUE=FALSE){ if(is.null(genenames) & !LISTRUE){ cdfname <- getCdfInfo(object) psets<- as.list(cdfname) psets<- psets[order(names(psets))] index <-unlist(sapply(psets, function(x) x[,2]),use.names=FALSE) return(exprs(object)[index,,drop=FALSE]) } else{ probes(object, "mm", genenames, LISTRUE=LISTRUE) } }) if( is.null(getGeneric("mm<-") )) setGeneric("mm<-", function(object, value) standardGeneric("mm<-")) setReplaceMethod("mm", "AffyBatch", function(object, value){ Dimnames <- dimnames(exprs(object)) cdfname <- getCdfInfo(object) psets<- as.list(cdfname) psets<- psets[order(names(psets))] mmIndex <-unlist(sapply(psets, function(x) x[,2]),use.names=FALSE) exprs(object)[mmIndex,] <- value dimnames(exprs(object)) <- Dimnames object }) ###probeset setGeneric("probeset", function(object, ...) standardGeneric("probeset")) setMethod("probeset", "AffyBatch", function(object, genenames=NULL, locations=NULL){ oldoptions <- getOption("BioC") if(is.null(locations)) ##use info in cdf envir <- getCdfInfo(object) else{ ##if the user gives a list of locations let them use that as enviromnet envir <- new.env() multiassign(names(locations), locations, envir) object@cdfName <- "envir" newoptions <- oldoptions newoptions$affy$probesloc[[1]]$what <- "environment" newoptions$affy$probesloc[[1]]$where <- parent.env(envir) options("BioC"=newoptions) } if(is.null(genenames)) genenames <- ls(envir) p.pps <- vector("list", length(genenames)) names(p.pps) <- genenames for (i in seq(along=genenames)) { i.pm <- indexProbes(object, "pm", genenames[i])[[1]] if (is.na(i.pm)[1]) intensity.pm <- matrix() else intensity.pm <- intensity(object)[i.pm, , drop=FALSE] i.mm <- indexProbes(object, "mm", genenames[i])[[1]] if (is.na(i.mm)[1]) intensity.mm <- matrix() else intensity.mm <- intensity(object)[i.mm, , drop=FALSE] p.pps[[i]] <- new("ProbeSet", id = genenames[i], pm = intensity.pm, mm = intensity.mm) } options("BioC"=oldoptions) return(p.pps) }) if (debug.affy123) cat("--->[[\n") ##[[: no more [[, because no more cel class # setMethod("[[", "AffyBatch", # function(x, i, j, ...) { ##no need for j # return(new("Cel", # intensity = matrix(intensity(x)[, i], ncol(x), nrow(x)), # name = sampleNames(x)[i], # cdfName = x@cdfName, # history = description(x)@preprocessing)) # }) ##[[ we need replacement that takes an entry by the Cel in value ##[ subseting. can only happen by sample. for now not by gene setMethod("[", "AffyBatch", function(x, i, j,..., drop=FALSE) { if (!missing(i) & missing(j)) { warning("The use of abatch[i,] and abatch[i] is deprecated. Please use abatch[,i] instead.\n") x <- x[,i] } if (!missing(j)) { phenoData(x) <- phenoData(x)[j, , ..., drop=FALSE] intensity(x) <- intensity(x)[ ,j, ..., drop=FALSE] if (!identical(se.exprs(x), new("matrix"))) { se.exprs(x) <- se.exprs(x)[ ,j, ..., drop=FALSE] } protocolData(x) <- protocolData(x)[j, , ..., drop=FALSE] } return(x) }) setReplaceMethod("[", "AffyBatch", function(x, i, j,..., value) { phenoData(x)[i,, ...] <- phenoData(value)[i, , ..., drop=FALSE] intensity(x)[,i] <- intensity(value)[ ,i,... , drop=FALSE] protocolData(x)[i,, ...] <- protocolData(value)[i, , ..., drop=FALSE] return(x) }) ## --- bg.correct if (debug.affy123) cat("--->bg.correct\n") if( is.null(getGeneric("bg.correct") )) setGeneric("bg.correct", function(object, method, ...) standardGeneric("bg.correct")) setMethod("bg.correct", signature(object="AffyBatch", method="character"), function(object, method=getOption("BioC")$affy$bgcorrect.method, ...) { ## simple for system to let one add background correction methods ## relies on naming convention method <- match.arg(method, bgcorrect.methods()) methodname <- paste("bg.correct.", method, sep="") if (! exists(methodname)) stop(paste("Unknown method (cannot find function", methodname, ")")) r <- do.call(methodname, alist(object, ...)) return(r) }) ## --- normalize.methods if( is.null(getGeneric("normalize.methods"))) setGeneric("normalize.methods", function(object) standardGeneric("normalize.methods")) setMethod("normalize.methods", signature(object="AffyBatch"), function(object) { normalize.AffyBatch.methods() }) ## ---normalize if (is.null(getGeneric("normalize"))) setGeneric("normalize", function(object, ...) standardGeneric("normalize")) setMethod("normalize", signature(object="AffyBatch"), function(object, method=getOption("BioC")$affy$normalize.method, ...) { method <- match.arg(method, normalize.AffyBatch.methods()) if (is.na(method)) stop("unknown method") method <- paste("normalize.AffyBatch", method, sep=".") object <- do.call(method, alist(object, ...)) ## collect info in the attribute "normalization" preproc <- c(description(object)@preprocessing, list(normalization = attr(object, "normalization"))) attr(object, "normalization") <- NULL ## and store it in MIAME MIAME <- description(object) MIAME@preprocessing <- preproc description(object) <- MIAME ## return(object) }) ## --- expression value computation if (debug.affy123) cat("--->computeExprSet\n") if( is.null(getGeneric("computeExprSet"))) setGeneric("computeExprSet", function(x, pmcorrect.method, summary.method, ...) standardGeneric("computeExprSet")) setMethod("computeExprSet", signature(x="AffyBatch", pmcorrect.method="character", summary.method="character"), function(x, pmcorrect.method, summary.method, ids=NULL, verbose=TRUE, summary.param=list(), pmcorrect.param=list()) { pmcorrect.method<- match.arg(pmcorrect.method, pmcorrect.methods()) summary.method <- match.arg(summary.method, express.summary.stat.methods()) ids <- unname(ids) n <- length(x) ## if 'ids' is NULL compute for all ids if (is.null(ids)) ids <- featureNames(x) m <- length(ids) pps.warnings <- vector("list", length=m) ## cheap trick to (try to) save time c.pps <- new("ProbeSet", pm=matrix(), mm=matrix()) ## matrix to hold expression values exp.mat <- matrix(NA, m, n) se.mat <- matrix(NA, m, n) if (verbose) { cat(m, "ids to be processed\n") countprogress <- 0 } ## loop over the ids mycall <- as.call(c(getMethod("express.summary.stat", signature=c("ProbeSet","character", "character")), list(c.pps, pmcorrect=pmcorrect.method, summary=summary.method, summary.param=summary.param, pmcorrect.param=pmcorrect.param)) ) ##only one character cause no more bg correct ##bg.correct=bg.method, param.bg.correct=bg.param, CDFINFO <- getCdfInfo(x) ##do it once! if (verbose) { pbt <- new("ProgressBarText", length(ids), barsteps = as.integer(20)) open(pbt) } for (i in seq(along=ids)) { if (verbose) { updateMe(pbt) } id <- ids[i] if (! exists(id, envir=CDFINFO)) { pps.warnings[[i]] <- paste("Unknown id", id) } else { ## locations for an id loc <- get(id, envir=CDFINFO) l.pm <- loc[, 1] if (ncol(loc) == 2) l.mm <- loc[ ,2] else l.mm <- integer() np <- length(l.pm) ##names are skipped c.pps@pm <- intensity(x)[l.pm, , drop=FALSE] c.pps@mm <- intensity(x)[l.mm, , drop=FALSE] ## generate expression values ## (wrapped in a sort of try/catch) mycall[[2]] <- c.pps ev <- try(eval(mycall), silent = TRUE) } if (! inherits(ev, "try-error")) { exp.mat[i, ] <- ev$exprs se.mat[i,] <- ev$se.exprs } else { pps.warnings[[i]] <- ev[1] } } if (verbose) { close(pbt) } dimnames(exp.mat) <- list(ids, sampleNames(x)) dimnames(se.mat) <- list(ids, sampleNames(x)) eset <- new("ExpressionSet", phenoData=phenoData(x), ## featureData picked up from exprs experimentData=experimentData(x), exprs=exp.mat, se.exprs=se.mat, annotation=annotation(x), protocolData=protocolData(x)) attr(eset, "pps.warnings") <- pps.warnings return(eset) }) ##some methods i was asked to add setMethod("image",signature(x="AffyBatch"), function(x, transfo=log, col=gray(c(0:64)/64), xlab="", ylab="",type=c("exprs","se.exprs"), main, ...){ scn <- prod(par("mfrow")) ask <- dev.interactive() which.plot <- 0 type <- match.arg(type) if (type == "se.exprs" && all(!dim(se.exprs(x)))){ stop("no se.exprs in object") } x.pos <- (1:nrow(x)) - (1 + getOption("BioC")$affy$xy.offset) y.pos <- (1:ncol(x)) - (1 + getOption("BioC")$affy$xy.offset) for(i in 1:length(sampleNames(x))){ which.plot <- which.plot+1; if(trunc((which.plot-1)/scn)==(which.plot-1)/scn && which.plot>1 && ask) par(ask=TRUE) if (type == "exprs"){ m <- exprs(x)[,i] } else { m <- se.exprs(x)[,i] } if (is.function(transfo)) { m <- transfo(m) } m <- as.matrix(rev(as.data.frame(matrix(m, nrow=length(x.pos), ncol=length(y.pos))))) if( missing(main) ){ main.cur=sampleNames(x)[i] } else { main.cur <- main } image(x.pos, y.pos, m, col=col, main=main.cur, xlab=xlab, ylab=ylab,,xaxt='n', yaxt='n', ...) par(ask=FALSE) } }) ##some special handling of main is needed setMethod("boxplot",signature(x="AffyBatch"), function(x, which="both", range=0, main, ...){ tmp <- description(x) if( missing(main) && (is(tmp, "MIAME")) ) main <- tmp@title tmp <- unlist(indexProbes(x,which)) tmp <- tmp[seq(1,length(tmp),len=5000)] boxplot(data.frame(log2(intensity(x)[tmp,])), main=main, range=range, ...) }) ###hist if (debug.affy123) cat("--->hist\n") if( is.null(getGeneric("hist")) ) setGeneric("hist") setMethod("hist",signature(x="AffyBatch"), function(x,...) plotDensity.AffyBatch(x,...)) if( is.null(getGeneric("mas5calls")) ) setGeneric("mas5calls", function(object,...) standardGeneric("mas5calls")) setMethod("mas5calls",signature(object="AffyBatch"), function(object,...) mas5calls.AffyBatch(object,...)) ##like for ExpressionSet "$.AffyBatch" <- function(affybatch, val) (pData(affybatch))[[as.character(val)]] affy/R/AffyRNAdeg.R0000644000175200017520000000576414516003533014767 0ustar00biocbuildbiocbuild"AffyRNAdeg" <- function (abatch,log.it=TRUE) { { data <- pm(abatch, LIST = TRUE) if(log.it==TRUE) data <- lapply(data,log2) names <- colnames(exprs(abatch)) probe.set.size <- function(x) { size <- dim(x)[1] return(size) } max.num <- sapply(data, probe.set.size) tab <- (table(max.num)) ord <- order(-as.numeric(tab)) K <- as.numeric(names(tab))[ord[1]] data <- data[max.num == K] } get.row <- function(x, i = 1) { return(x[i, ]) } get.col <- function(x, i = 1) { return(x[, i]) } rowstack <- function(x, i = 1) { return(t(sapply(x, get.row, i))) } colstack <- function(x, i = 1) { return(t(sapply(x, get.col, i))) } N <- length(data) n <- dim(data[[1]])[2] mns <- matrix(nrow = n, ncol = K) sds <- mns for (i in 1:K) { data.stack <- rowstack(data, i) if(dim(data[[1]])[2]==1) data.stack <- t(data.stack) mns[, i] <- colMeans(data.stack) sds[, i] <- apply(data.stack, 2, sd) } mns.orig <- mns mn <- mns[, 1] mns <- sweep(mns, 1, mn) mns <- mns/(sds/sqrt(N)) lm.stats <- function(x) { index <- 0:(length(x) - 1) ans <- summary(lm(x ~ index))$coefficients[2, c(1, 4)] return(ans) } stats <- apply(mns, 1, lm.stats) answer <- list(N, names, mns.orig, sds/sqrt(N), stats[1, ], stats[2, ]) names(answer) <- c("N", "sample.names", "means.by.number", "ses", "slope", "pvalue") return(answer) } "summaryAffyRNAdeg" <- function (rna.deg.obj, signif.digits = 3) { temp.table <- rbind(signif(rna.deg.obj$slope, signif.digits), signif(rna.deg.obj$pvalue, signif.digits)) colnames(temp.table) <- rna.deg.obj$sample.names rownames(temp.table) <- c("slope", "pvalue") ##write.table(temp.table, file = "", quote = FALSE) return(temp.table) } "plotAffyRNAdeg" <- function (rna.deg.obj,transform="shift.scale",cols=NULL, ...) { if(!is.element(transform,c("shift.scale","shift.only","neither"))) stop("Tranform must be 'shift.scale','shift.only', or 'neither'") mns <- rna.deg.obj$means.by.number if(is.null(cols)) cols=rep(4,dim(mns)[1]) ylab="Mean Intensity" if(transform=="shift.scale"){ sds <- rna.deg.obj$ses mn <- mns[, 1] mns <- sweep(mns, 1, mn) mns <- mns/(sds) mns <- sweep(mns, 1, 1:(dim(mns)[1]), "+") ylab <- paste(ylab,": shifted and scaled") }else if(transform=="shift.only"){ mn <- mns[, 1] mns <- sweep(mns, 1, mn) mns <- sweep(mns, 1, 1:(dim(mns)[1]), "+") ylab <- paste(ylab,": shifted") } plot(-2, -1, pch = "", xlim = range(-1, (dim(mns)[2])), ylim = range(min(as.vector(mns)) - 1, max(as.vector(mns)) + 1), xlab = "5' <-----> 3'\n Probe Number ", ylab = ylab, axes = FALSE, main = "RNA degradation plot", ...) axis(1) axis(2) for (i in 1:dim(mns)[1]) lines(0:((dim(mns)[2]-1)), mns[i, ],col=cols[i]) } affy/R/ProbeSet.R0000644000175200017520000001142414516003533014572 0ustar00biocbuildbiocbuild ## A ProbeSet holds probe values for a probe pairs set(*) accross a batch of experiments. ## methods 'express.summary.stat' returns of expression value per experiement in the ## batch, and 'bg.correct' does background correction (in some sense... the MM probes ## were created to measure unspecific hybridization. People thought that doing ## PM - MM would remove background noise. The method 'bg.correct' accepts extra parameters ## through '...' (can be used to pass background correction parameters common to different ## ProbeSet) ## ## - ## (*) : a probe pair set is the set of probes pairs(**) related to an affyid. Generally a ## a probe pair set has 20 elements. ## (**): a probe pair (or atom) is a pair of PM/MM values ## if (debug.affy123) cat("-->initProbeSet\n") setClass("ProbeSet", representation(id="character", pm="matrix", mm="matrix"), prototype=list(pm=matrix(), mm=matrix())) setMethod("show", "ProbeSet", function(object) { cat("ProbeSet object:\n") cat(" id=", object@id, "\n", sep="") cat(" pm=", nrow(object@pm), "probes x ", ncol(object@pm), " chips\n") }) ##DEBUG: what to do with that ? ## --> with what ? setMethod("sampleNames", "ProbeSet", function(object) colnames(object)) setMethod("colnames", signature(x="ProbeSet"), function(x ,do.NULL=FALSE, prefix="row") { cnames<-colnames(pm(x)) if (is.null(cnames)) { if (do.NULL) { warning("No column names for ProbeSet") } else { cnames <- paste(prefix, 1:ncol(x@pm)) } } return(cnames) }) ## pm if( is.null(getGeneric("pm"))) setGeneric("pm", function(object) standardGeneric("pm")) setMethod("pm", "ProbeSet", function(object) object@pm) if( is.null(getGeneric("pm<-"))) setGeneric("pm<-", function(object, value) standardGeneric("pm<-")) setReplaceMethod("pm", signature=c("ProbeSet", "matrix"), function(object, value) { if (! all(dim(value) == dim(object@mm))) stop("dimension mismatch between 'pm' and 'mm'") object@pm <- value }) ## mm if( is.null(getGeneric("mm"))) setGeneric("mm", function(object) standardGeneric("mm")) setMethod("mm", "ProbeSet", function(object) object@mm) if( is.null(getGeneric("mm<-"))) setGeneric("mm<-", function(object, value) standardGeneric("mm<-")) setReplaceMethod("mm", signature=c("ProbeSet", "matrix"), function(object, value) { if (sum(dim(value) == dim(object@mm)) != 2) stop("dimension mismatch between 'pm' and 'mm'") object@mm <- value }) ## method express.summary.stat if( is.null(getGeneric("express.summary.stat"))) setGeneric("express.summary.stat", function(x, pmcorrect, summary, ...) standardGeneric("express.summary.stat")) setMethod("express.summary.stat",signature(x="ProbeSet", pmcorrect="character", summary="character"), function(x, pmcorrect, summary, summary.param=list(), pmcorrect.param=list()) { pmcorrect <- match.arg(pmcorrect, pmcorrect.methods()) summary <- match.arg(summary, express.summary.stat.methods()) ## simple for system to let one add background correction methods ## relies on naming convention pmcorrect.methodname <- paste("pmcorrect.", pmcorrect, sep="") summary.methodname <- paste("generateExprVal.method.", summary, sep="") if (! exists(summary.methodname)) stop(paste("Unknown method (cannot find function", summary.methodname, ")")) if (! exists(pmcorrect.methodname)) stop(paste("Unknown method (cannot find function", pmcorrect.methodname, ")")) ## NOTE: this could change... #m <- do.call(bg.correct, c(alist(x@pm, x@mm), param.bg.correct)) pm.corrected <- do.call(pmcorrect.methodname, c(alist(x), pmcorrect.param)) r <- do.call(summary.methodname, c(alist(pm.corrected), summary.param)) ##DEBUG: name stuff to sort #names(r) <- names(allprobes) return(r) }) setMethod("barplot",signature(height="ProbeSet"),function(height,...) barplot.ProbeSet(height,...)) if( is.null(getGeneric("mas5calls")) ) setGeneric("mas5calls", function(object,...) standardGeneric("mas5calls")) setMethod("mas5calls",signature(object="ProbeSet"), function(object,...) mas5calls.ProbeSet(object,...)) affy/R/ProgressBarText.R0000644000175200017520000000610614516003533016146 0ustar00biocbuildbiocbuilddebug.affy123 <- FALSE setClass("ProgressBarText", representation(steps = "integer", barsteps = "integer", internals = "environment")) setMethod("initialize", "ProgressBarText", function(.Object, steps, barsteps = 10, internals = NULL) { ##.Object <- callNextMethod() if ( ! is.null(internals)) { stop("slot 'internals' is for internal use !") } .Object@barsteps = barsteps .Object@internals = new.env() assign("milestones.i", as.integer(1), envir=.Object@internals) assign("increment", as.integer(1), envir=.Object@internals) assign("milestones", as.integer(seq(1, steps, length=barsteps)), envir=.Object@internals) assign("i", as.integer(0), envir=.Object@internals) return(.Object) }) setMethod("open", "ProgressBarText", function(con, header = TRUE) { if (header) { cat("|", paste(rep(" ", con@barsteps), collapse=""), "|\n", sep="") } cat("|") increment <- get("increment", con@internals) milestones.i <- get("milestones.i", con@internals) milestones <- get("milestones", con@internals) while(milestones.i > length(milestones)) { cat("#") } if (.Platform$OS.type == "windows") flush.console() }) ## to avoid 'loosing' the default update. ## (not sure this is the most elegant way to do this) setGeneric("updateMe", function(object, ...) standardGeneric("updateMe")) setMethod("updateMe", "ProgressBarText", function(object) { increment <- get("increment", object@internals) i <- get("i", object@internals) + increment milestones.i <- get("milestones.i", object@internals) milestones <- get("milestones", object@internals) touched <- FALSE while(milestones.i <= length(milestones) && i >= milestones[milestones.i]) { cat("#") milestones.i <- milestones.i + increment touched <- TRUE } ## the 'touch' thing appears to make it save 0.1 sec / 100000 iteration ## (which makes it absolutely mandatory :) ). if (touched) { assign("milestones.i", milestones.i, envir = object@internals) if (.Platform$OS.type == "windows") flush.console() } assign("i", i, , envir = object@internals) }) setMethod("close", "ProgressBarText", function(con) { increment <- get("increment", con@internals) milestones.i <- get("milestones.i", con@internals) milestones <- get("milestones", con@internals) while(milestones.i <= length(milestones)) { cat("#") milestones.i <- milestones.i + increment } assign("milestones.i", milestones.i, envir = con@internals) cat("|\n") if (.Platform$OS.type == "windows") flush.console() }) affy/R/avdiff.R0000644000175200017520000000070014516003533014301 0ustar00biocbuildbiocbuildavdiff <- function(x,verbose=FALSE){ if(missing(x)) stop("Argument x missing, with no default\n") cat("Computing average difference for",dim(x$pm)[2],"columns") avdiff <- apply(x$pm-x$mm,2,function(y){ cat(".") tapply(y,x$name,function(z){ o <- order(z) zz <- z[-c(o[1],o[length(z)])] #take out biggest and smallest mean(z[abs(z-mean(zz))<3*sd(zz)]) }) }) colnames(avdiff) <- x$chip.names return(avdiff) } affy/R/barplot.ProbeSet.R0000644000175200017520000000204114516003533016227 0ustar00biocbuildbiocbuildbarplot.ProbeSet <- function(height, xlab="Probe pair",ylab="Intensity", main=NA, col.pm="red", col.mm="blue", beside=TRUE, names.arg="pp", ask = TRUE, scale = TRUE, ...) { opar <- par()$ask par(ask=ask) on.exit(par(ask=opar)) if (names.arg == "pp") { names.arg <- seq(1, nrow(pm(height))) } col <- c(col.pm, col.mm) if (scale) { ylim <- range(c(pm(height), mm(height)), na.rm=TRUE) } else { ylim <- NULL } if (is.na(main)) { main <- paste(height@id, "( sample", 1:ncol(pm(height)), ")") } else { main <- rep(main, length=ncol(pm(height))) } for (i in 1:ncol(pm(height))) { hh <- rbind(pm(height)[, i], mm(height)[, i]) barplot(hh, xlab=xlab, ylab=ylab, main=main[i], col=col, beside=beside, names.arg=names.arg, ylim = ylim, ...) } } affy/R/bg.Affy.chipwide.R0000644000175200017520000000266514516003533016125 0ustar00biocbuildbiocbuildbg.correct.mas <- function(object, griddim=16) { nchips <- length(object) pm.index <- unique(unlist(indexProbes(object, "pm"))) mm.index <- unique(unlist(indexProbes(object, "mm"))) ## some chips have some probesets without MM probes ## which will return an NA in mm.index mm.index <- mm.index[!is.na(mm.index)] rows <- nrow(object) cols <- ncol(object) allintensities <- intensity(object)[c(pm.index, mm.index), ] # note that the indexing is +1 more than you'd expect because # the c code expects it that way ## (note about the remark above: R indexing starts at 1 and not at 0, ## that's why the indexing is done this way. The package is primarily done to ## be used with R...) allx <- c(pm.index-1, mm.index-1) %% nrow(object) +1 ally <- c(pm.index-1, mm.index-1) %/% nrow(object) + 1 nprobes <- length(allx) corrected <- matrix(.C("affy_background_adjust_R", as.double(as.vector(allintensities)), as.integer(allx), as.integer(ally), as.integer(nprobes), as.integer(nchips), as.integer(rows), as.integer(cols), as.integer(griddim), PACKAGE="affy")[[1]], nprobes, nchips) intensity(object)[c(pm.index, mm.index), ] <- corrected ## and what with the 'non pm or mm' probes ? ## answer: they are not used per Affymetrix Statistical Algorithms Description Document. return(object) } affy/R/bg.R0000644000175200017520000000333514516003533013441 0ustar00biocbuildbiocbuild####These functions take an AffyBatch object "background correct" ####the pms and return an AffyBatch with the background corrected PMs ### bg.parameters <- function(pm, n.pts=2^14){ max.density <- function(x, n.pts){ aux <- density(x, kernel="epanechnikov", n=n.pts, na.rm=TRUE) aux$x[order(-aux$y)[1]] } pmbg <- max.density(pm,n.pts) ##Log helps detect mode bg.data <- pm[pm < pmbg] ##do it again to really get the mode pmbg <- max.density(bg.data,n.pts) bg.data <- pm[pm < pmbg] bg.data <- bg.data - pmbg bgsd <- sqrt(sum(bg.data^2)/(length(bg.data)-1))*sqrt(2)#/.85 sig.data <- pm[pm > pmbg] sig.data <- sig.data-pmbg expmean <- max.density(sig.data,n.pts) alpha <- 1/expmean mubg <- pmbg list(alpha=alpha,mu=mubg,sigma=bgsd) } bg.adjust <- function(pm, n.pts=2^14, ...){ param <- bg.parameters(pm,n.pts) b <- param$sigma pm <- pm - param$mu - param$alpha*b^2 pm + b*((1./sqrt(2*pi))*exp((-1./2.)*((pm/b)^2)))/pnorm(pm/b) } bg.correct.none <- function(object, ...) object ##bg.correct.subtractmm <- function(object){ ## pm(object) <- pm(object) - mm(object) ## return(object) ##} ###bg.correct.rma <- function(object, ...){ ### pm(object) <- apply(pm(object),2,bg.adjust) ### return(object) ##} ## ## this function calls the c code as an alternative to the R code above. ## it should help end the disagreement between rma() and expresso() ## bg.correct.rma <- function(object,...){ pm(object) <- rma.background.correct(pm(object),copy=FALSE) return(object) } ## --- pmcorrect things pmcorrect.subtractmm <- function(object){ pm.corrected <- pm(object) - mm(object) return(pm.corrected) } pmcorrect.pmonly <- function(object) { return(pm(object)) } affy/R/expresso.R0000644000175200017520000000764514516003533014731 0ustar00biocbuildbiocbuildexpresso <- function(afbatch, ## -- bg.correct=TRUE, bgcorrect.method = NULL, bgcorrect.param = list(), ## -- normalize = TRUE, normalize.method = NULL, normalize.param=list(), ## -- pmcorrect.method = NULL, pmcorrect.param = list(), ## -- summary.method = NULL, summary.param = list(), summary.subset = NULL, ## --- verbose = TRUE, widget = FALSE ) { # JZ added this function setCorrections <- function(){ bioc.opt <- getOption("BioC") if(bg.correct){ if(is.null(bgcorrect.method)){ BGMethods <- bgcorrect.methods() }else{ BGMethods <- bgcorrect.method } }else{ BGMethods <- "None" } if(normalize){ if(is.null(normalize.method)){ normMethods <- normalize.methods(afbatch) }else{ normMethods <- normalize.method } }else{ normMethods <- "None" } # Default for this one may not be correct if(is.null(pmcorrect.method)){ PMMethods <- pmcorrect.methods() }else{ PMMethods <- pmcorrect.method } # Default for this one may not be correct if(is.null(summary.method)){ expMethods <- generateExprSet.methods() }else{ expMethods <- summary.method } corrections <- expressoWidget(BGMethods, normMethods, PMMethods, expMethods, bioc.opt$affy$bgcorrect.method, bioc.opt$affy$normalize.method, bioc.opt$affy$pmcorrect.method, bioc.opt$affy$summary.method) if(!is.null(corrections)){ if(corrections[["BG"]] != "None"){ bgcorrect.method <<- corrections[["BG"]] } if(corrections[["NORM"]] != "None"){ normalize.method <<- corrections[["NORM"]] } if(corrections[["PM"]] != "None"){ pmcorrect.method <<- corrections[["PM"]] } if(corrections[["EXP"]] != "None"){ summary.method <<- corrections[["EXP"]] } }else{ stop("Aborted by user") } } if (widget) { requireNamespace("tkWidgets") } nchips <- length(afbatch) ###background stuff must be added before normalization! if(widget){ setCorrections() } ## -- summary of what will be done if (verbose) { if (bg.correct){ cat("background correction:", bgcorrect.method, "\n") } if (normalize) { cat("normalization:", normalize.method, "\n") } cat("PM/MM correction :", pmcorrect.method, "\n") cat("expression values:", summary.method, "\n") } ## -- background correct (if needed) if (bg.correct) { if (verbose) cat("background correcting...") afbatch <- do.call("bg.correct", c(alist(afbatch, method=bgcorrect.method), bgcorrect.param)) if (verbose) cat("done.\n") } ## -- normalize (if wished) if (normalize) { if (verbose) cat("normalizing...") afbatch <- do.call(BiocGenerics::normalize, c(alist(afbatch, normalize.method), normalize.param)) if (verbose) cat("done.\n") } eset <- computeExprSet(afbatch, summary.method=summary.method, pmcorrect.method= pmcorrect.method, ids=summary.subset, summary.param=summary.param, pmcorrect.param=pmcorrect.param) return(eset) } affy/R/expressoWidget.R0000644000175200017520000001040114516003533016055 0ustar00biocbuildbiocbuild# A function that takes user inputs for correction methods for # expresso (affy). Default values can be missing, in which case the # first element will be chosen as the default. expressoWidget <- function(BGMethods, normMethods, PMMethods, expMethods, BGDefault, normDefault, PMDefault, expDefault){ methodList <- list() END <- FALSE if(any(missing(BGMethods), missing(normMethods), missing(PMMethods), missing(expMethods))){ stop("At least one of the method arguments is missing") } if(any(c(length(BGMethods), length(normMethods), length(PMMethods), length(expMethods)) == 0)){ stop("At least one of the method argument is of length 1") } if(missing(BGDefault)){ BGM <- tcltk::tclVar(BGMethods[1]) }else{ BGM <- tcltk::tclVar(BGDefault) } if(missing(normDefault)){ NMM <- tcltk::tclVar(normMethods[1]) }else{ NMM <- tcltk::tclVar(normDefault) } if(missing(PMDefault)){ PMM <- tcltk::tclVar(PMMethods[1]) }else{ PMM <- tcltk::tclVar(PMDefault) } if(missing(expDefault)){ EXM <- tcltk::tclVar(expMethods[1]) }else{ EXM <- tcltk::tclVar(expDefault) } quit <- function(){ tcltk::tkdestroy(base) } end <- function(){ END <<- TRUE methodList[["BG"]] <<- tcltk::tclvalue(BGM) methodList[["NORM"]] <<- tcltk::tclvalue(NMM) methodList[["PM"]] <<- tcltk::tclvalue(PMM) methodList[["EXP"]] <<- tcltk::tclvalue(EXM) quit() } base <- tcltk::tktoplevel() ## post -- hook on.exit(tcltk::tkdestroy(base)) tcltk::tktitle(base) <- "Expresso methods selection" ## Description text tcltk::tkpack(tcltk::tklabel(base, text = "Welcome to Expresso methods selection"), expand = FALSE, fill = "x", padx = 5, pady = 5) tcltk::tkpack(tcltk::tklabel(base, text = paste("You need to choose correction", "methods or go with the defaults")), expand = FALSE, fill = "x", padx = 5) ## Selections for correction methods methodFrame <- tcltk::tkframe(base) ## Background selection BGLabel <- tcltk::tklabel(methodFrame, text = "Background correction") BGDropdown <- tcltk::tkframe(methodFrame) widgetTools::dropdownList(BGDropdown, BGMethods, BGM, 20, tcltk::tclvalue(BGM), TRUE) tcltk::tkgrid(BGLabel, BGDropdown) tcltk::tkgrid.configure(BGLabel, sticky = "e") tcltk::tkgrid.configure(BGDropdown, sticky = "w") ## Normlization NMLabel <- tcltk::tklabel(methodFrame, text = "Normalization") NMDropdown <- tcltk::tkframe(methodFrame) widgetTools::dropdownList(NMDropdown,normMethods, NMM, 20, tcltk::tclvalue(NMM), TRUE) tcltk::tkgrid(NMLabel, NMDropdown) tcltk::tkgrid.configure(NMLabel, sticky = "e") tcltk::tkgrid.configure(NMDropdown, sticky = "w") ## PM correction PMLabel <- tcltk::tklabel(methodFrame, text = "PM correction") PMDropdown <- tcltk::tkframe(methodFrame) widgetTools::dropdownList(PMDropdown, PMMethods, PMM, 20, tcltk::tclvalue(PMM), TRUE) tcltk::tkgrid(PMLabel, PMDropdown) tcltk::tkgrid.configure(PMLabel, sticky = "e") tcltk::tkgrid.configure(PMDropdown, sticky = "w") ## PM correction EXLabel <- tcltk::tklabel(methodFrame, text = "Expression") EXDropdown <- tcltk::tkframe(methodFrame) widgetTools::dropdownList(EXDropdown, expMethods, EXM, 20, tcltk::tclvalue(EXM), TRUE) tcltk::tkgrid(EXLabel, EXDropdown) tcltk::tkgrid.configure(EXLabel, sticky = "e") tcltk::tkgrid.configure(EXDropdown, sticky = "w") tcltk::tkpack(methodFrame, expand = TRUE, fill = "both", padx = 5, pady = 10) butFrame <- tcltk::tkframe(base) quitBut <- tcltk::tkbutton(butFrame, text = "Quit", width = 7, command = quit) endBut <- tcltk::tkbutton(butFrame, text = "Select", width = 7, command = end) tcltk::tkgrid(quitBut, endBut, padx = 5) tcltk::tkpack(butFrame, expand = FALSE, fill = "x", pady = 5) tcltk::tkwait.window(base) if(END){ return(methodList) }else{ return(NULL) } } affy/R/fit.li.wong.R0000644000175200017520000002045414516003533015210 0ustar00biocbuildbiocbuildfit.li.wong <- function(data.matrix, remove.outliers=TRUE, normal.array.quantile=0.5, normal.resid.quantile=0.9, large.threshold=3, large.variation=0.8, outlier.fraction=0.14, delta = 1e-06,maxit=50,outer.maxit=50, verbose=FALSE, ...){ if(missing(data.matrix)) stop("Argument data.matrix missing, with no default") II <- dim(data.matrix)[1] ##II instrad of I cause I is a fuction in R J <- dim(data.matrix)[2] if(J==1){ warning("Li and Wong's algorithm is not suitable when only one probe pair") return(list(theta = as.vector(data.matrix), phi = 1, sigma.eps = NA, sigma.theta = NA, sigma.phi=NA, theta.outliers=NA, phi.outliers=NA, single.outliers=NA,convergence1=NA,convergence2=NA,iter = NA, delta = NA)) } cI <- II ##current I cJ <- J ##current J theta.outliers.old <- rep(FALSE, II) ##ith entry will be true if theta_i is an outlier phi.outliers.old <- rep(FALSE, J) ##jth entry will be true if phi_j is an outlier single.outliers.old <- matrix(FALSE, II, J) ##ij entry will be true if y_is an outlier theta.outliers <- theta.outliers.old ##need this to now if change ocurred in outliers phi.outliers <- phi.outliers.old ##need this to know if chages occured in outlies single.outliers <- single.outliers.old flag1 <- NA ##these will be false if convergence not reacher, flag2 <- NA ## this will be false if outliers respectively cuase iter to stop if(remove.outliers){ flag1 <- TRUE; flag2<-TRUE original.data.matrix <- data.matrix ##so we can get it back after outlier removal change.theta <- 1 #start with 1 change.phi <- 1 change.single <- 1 outer.iter <- 0 while(flag1 & flag2 & change.theta+change.phi+change.single >0 & outer.iter < outer.maxit) { outer.iter <- outer.iter + 1 if((outer.iter%%3==0 & change.theta>0) | (outer.iter%%3==1 & change.phi>0)){ #something has to change ##starting values phi <- colMeans(data.matrix) c <- sqrt(cJ/sum(phi[!phi.outliers]^2)) phi <- c * phi theta <- (data.matrix[, !phi.outliers, drop=FALSE] %*% phi[!phi.outliers, drop=FALSE])/cJ iter <- 0 change <- 1 #start with one theta.old <- rep(0, II) while(change > delta & iter < maxit) { iter <- iter + 1 phi <- t(data.matrix[!theta.outliers, ,drop=FALSE]) %*% theta[!theta.outliers, drop=FALSE] ##ignore the outliers c <- sqrt(cJ/sum(phi[!phi.outliers, drop=FALSE]^2)) phi <- c * phi theta <- (data.matrix[,!phi.outliers, drop=FALSE] %*% phi[!phi.outliers, drop=FALSE])/cJ change <- max(abs(theta[!theta.outliers] - theta.old[!theta.outliers])) if(verbose) cat(paste("Outlier iteration:",outer.iter,"estimation iteration:",iter,"chage=",change,"\n")) theta.old <- theta } if(iter>=maxit){ ##convergence not reached. might as well get out warning(paste("No convergence in inner loop after",iter,"in outerler tieration",outer.iter,"\n")) flag1 <- FALSE } if(mean(phi[!phi.outliers]<0)>.5){ ##for identifiability.. theta*phi = (-theta)*(-phi), i require that most phis are positive theta <- -theta phi <- -phi } theta <- as.vector(theta) phi <- as.vector(phi) data.matrixhat <- outer(theta, phi) resid <- data.matrix-data.matrixhat } ##DEALING WITH OUTLIERS ##we alternate removal of outliers ##if even iteration take out thetas that are outliers (as defined by Li and Wong). if(outer.iter%%3==1){ ## we start with single outliers single.outliers <- resid > large.threshold*quantile(abs(resid),normal.resid.quantile) single.outliers[rowSums(single.outliers) > outlier.fraction*cJ,]<-rep(FALSE,J) ##probably chip oulier, defer calling outlier single.outliers[,colSums(single.outliers) > outlier.fraction*cI]<-rep(FALSE,II) ##probably probe outlier, defer calling outlier data.matrix[single.outliers] <- data.matrixhat[single.outliers] data.matrix[!single.outliers] <- original.data.matrix[!single.outliers] change.single <- sum(abs(single.outliers.old-single.outliers)) #sum will be total of changes single.outliers.old <- single.outliers } else{ sigma.theta <- sqrt(rowSums(resid[, !phi.outliers, drop=FALSE]^2)/(cJ - 1)) sigma.phi <- sqrt(colSums(resid[!theta.outliers, , drop=FALSE]^2)/(cI - 1)) ###THETA OUTLIERS if(outer.iter%%3==2){ theta.outliers <- sigma.theta > large.threshold*quantile(sigma.theta,normal.array.quantile) | theta^2/sum(theta^2) > large.variation cI <- sum(!theta.outliers) if(cI<3) { warning("No convergence achieved, too many outliers") flag2 <- FALSE } ##single outliers in outlier chips are not longer single outliers single.outliers[theta.outliers,] <- rep(FALSE,J) data.matrix[single.outliers] <- data.matrixhat[single.outliers] data.matrix[!single.outliers]<-original.data.matrix[!single.outliers] change.theta <- sum(abs(theta.outliers.old-theta.outliers)) #sum will be total of changes change.single <- sum(abs(single.outliers.old-single.outliers)) #sum will be total of changes theta.outliers.old <- theta.outliers } ##PHI OUTLIERS else{ phi.outliers <- sigma.phi > large.threshold*quantile(sigma.phi,normal.array.quantile) | phi^2/sum(phi^2) > large.variation | phi <0 cJ <- sum(!phi.outliers) if(cJ<3) { warning("No convergence achieved, too many outliers") flag2 <- FALSE } single.outliers[,phi.outliers] <- rep(FALSE,II) data.matrix[single.outliers] <- data.matrixhat[single.outliers] data.matrix[!single.outliers]<-original.data.matrix[!single.outliers] change.phi <- sum(abs(phi.outliers.old-phi.outliers)) change.single <- sum(abs(single.outliers.old-single.outliers)) #sum will be total of changes phi.outliers.old <- phi.outliers } } if(verbose){ cat("chips used=",cI,", probes used=",cJ,", single outler=",sum(single.outliers),"\n") cat("Number of changes: single=",change.single,", theta=",change.theta,", phi=",change.phi,"\n",sep="") } } if(outer.iter>=outer.maxit){ warning("No convergence achieved in outlier loop\n") flag2 <- FALSE } all.outliers <- outer(theta.outliers,phi.outliers,FUN="|") | single.outliers sigma <- sqrt(sum(resid[!all.outliers]^2)/sum(!all.outliers)) ##in case we leave iteration and these havent been defined sigma.theta <- sqrt(rowSums(resid[,!phi.outliers, drop=FALSE]^2)/(cJ - 1)) sigma.phi <- sqrt(colSums(resid[!theta.outliers, ,drop=FALSE]^2)/(cI - 1)) } ###code for NO OUTLIER REMOVAL else{ flag1 <- TRUE phi <- colMeans(data.matrix) c <- sqrt(J/sum(phi^2)) phi <- c * phi theta <- (data.matrix %*% phi)/J iter <- 0 change <- 1 theta.old <- rep(0, II) while(change > delta & iter < maxit) { iter <- iter + 1 phi <- t(data.matrix) %*% theta c <- sqrt(J/sum(phi^2)) phi <- c * phi theta <- (data.matrix %*% phi)/J change <- max(abs(theta - theta.old)) if(verbose) cat(paste("Iteration:",iter,"chage=",change,"\n")) theta.old <- theta } if(iter>=maxit){ warning(paste("No convergence after",iter,"iterations.\n")) flag1 <- FALSE } if(mean(phi[!phi.outliers]<0)>.5){ ##for identifiability.. theta*phi = (-theta)*(-phi), i require that most phis are positive theta <- -theta phi <- -phi } theta <- as.vector(theta) phi <- as.vector(phi) data.matrixhat <- outer(theta, phi) sigma.theta <- sqrt(rowSums((data.matrix - data.matrixhat)^2)/(J - 1)) sigma.phi <- sqrt(colSums((data.matrix - data.matrixhat)^2)/(II - 1)) sigma <- sqrt(sum((data.matrix - data.matrixhat)^2)/(II * J)) } return(list(theta = theta, phi = phi, sigma.eps = sigma, sigma.theta = sigma.theta, sigma.phi=sigma.phi,theta.outliers=theta.outliers,phi.outliers=phi.outliers,single.outliers=single.outliers,convergence1=flag1,convergence2=flag2,iter = iter, delta = change)) } affy/R/generateExprVal.method.avgdiff.R0000644000175200017520000000051114516003533021022 0ustar00biocbuildbiocbuild## Currently, the input is a 2 matrices a pm and a mm ##avdiff is more like median than mean, it would be nice to actually have ##avfif ##added typical se of the mean as returned se generateExprVal.method.avgdiff <- function(probes, ...) { list(exprs=apply(probes, 2, median),se.exprs=apply(probes,2,sd)/sqrt(nrow(probes))) } affy/R/generateExprVal.method.liwong.R0000644000175200017520000000052714516003533020722 0ustar00biocbuildbiocbuildgenerateExprVal.method.liwong <- function(probes, ...) { probes <- t(probes) if (ncol(probes) == 1) { warning("method liwong unsuitable when only one probe pair") list(exprs=as.vector(probes),se.exprs=rep(NA,length(probes))) } else { tmp <- fit.li.wong(probes, ...) list(exprs=tmp$theta,se.exprs=tmp$sigma.theta) } } affy/R/generateExprVal.method.mas.R0000644000175200017520000000127214516003533020201 0ustar00biocbuildbiocbuildgenerateExprVal.method.mas <- function(probes, ...) { probes <- log2(probes) M <- ncol(probes) slg <- rep(NA,M) for (i in 1:ncol(probes)) { slg[i] <- tukey.biweight(probes[ ,i], ...) } return(list(exprs=2^slg,se.exprs=rep(NA,M))) } affy.scalevalue.exprSet <- function(eset, sc=500, analysis="absolute") { analysis <- match(analysis, c("absolute", "comparison")) if(analysis == 1) nf <- 1 else stop("sorry! comparison not implemented.") for (i in 1:ncol(exprs(eset))) { slg <- exprs(eset)[, i] sf <- sc / mean(slg, trim=0.02) reported.value <- nf * sf * slg exprs(eset)[, i] <- reported.value } return(eset) } affy/R/generateExprVal.method.medianpolish.R0000644000175200017520000000013314516003533022070 0ustar00biocbuildbiocbuildgenerateExprVal.method.medianpolish <- function(probes, ...) medianpolish(probes, ...) affy/R/generateExprVal.method.playerout.R0000644000175200017520000000217014516003533021443 0ustar00biocbuildbiocbuildgenerateExprVal.method.playerout <- function(probes, weights=FALSE, optim.method="L-BFGS-B"){ probes <- t(probes) nprobes <- ncol(probes) ## skip if only one probe if (nprobes == 1) return(t(probes)) ## I do not know to which extend the use of optim ## is really equivalent to the use of nlminb in S-plus S1 <- optim(runif(nprobes), playerout.costfunction, method=optim.method, control=list(maxit=500), y=probes) ##S1 <- nlm(playerout,runif(20),iterlim=500,y=t(y)) r <- c(probes %*% S1$par / sum(S1$par)) if (weights) attr(r,"weights") <- S1$par return(list(exprs=r,se.exprs=rep(NA,length(r)))) } ## The loss function: playerout.costfunction <- function(w, y) { N <- length(w) # Number of players J <- length(y)/N # Number of games (the number of games is the number of chips used) sumw <- sum(w) tx <- y %*% w # Full weighted score at each game pl <- matrix(0,J,N) # Loss at each game due to each player for(j in 1:J) pl[j,] <- w * y[j,] - (tx[j] - w * y[j,]) / (sumw - w) sum(pl^2) # Loss } affy/R/getCDFenv.R0000644000175200017520000001303614516003533014655 0ustar00biocbuildbiocbuildgetCdfInfo <- function(object, how=getOption("BioC")$affy$probesloc, verbose=FALSE) { ## cdfname is the cdf environment ## methods is a vector detailing how to get the file - one of ## 'library', 'bioC' if (length(how) == 0) stop("No available method to obtain CDF file") cdfname <- cdfName(object) badOut <- list() for (i in 1:length(how)) { cur <- how[[i]] out <- switch(cur$what, "environment" = cdfFromEnvironment(cdfname, cur$where, verbose), "libPath" = cdfFromLibPath(cdfname, cur$where, verbose=verbose), "bioC" = cdfFromBioC(cdfname, cur$where, verbose) ) if (is.environment(out)) return(out) else badOut <- c(badOut, out) } stop(paste("Could not obtain CDF environment, problems encountered:", paste(unlist(badOut),collapse="\n"),sep="\n")) } cdfFromEnvironment <- function(cdfname, where, verbose=TRUE) { if (verbose) print(paste("Attempting to locate",cdfname,"in specified environment")) if (exists(cdfname, inherits=FALSE, where=where)) return(as.environment(get(cdfname,inherits=FALSE,envir=where))) else { if (verbose) print(paste("Specified environment does not contain",cdfname)) return(list(paste("Specified environment does not contain",cdfname))) } } cdfFromBioC <- function(cdfname, lib=.libPaths()[1], verbose=TRUE) { cdfname <- cleancdfname(cdfname) if (verbose) print(paste("Attempting to obtain",cdfname,"from Bioconductor website")) ## First try libPaths libs <- .libPaths() if (!all(lib %in% libs)) libs <- unique(c(lib, libs)) result <- cdfFromLibPath(cdfname, lib=libs, verbose=verbose) if (is.environment(result)) return(result) if (length(lib) > 1) { warning("Ignoring all but first element of argument lib") lib <- lib[1] } if (verbose) print(paste("The environment ",cdfname," was not found in", " these directories: ", paste(libs, collapse=", "), ". Now searching the internet repository.", sep="")) if (verbose) print(paste("Checking to see if your internet connection works ...")) if (testBioCConnection()) { ## Check for file permissions if (file.access(lib, mode=0) < 0) { if (verbose) { print(paste("Directory",lib,"does not seem to exist.\n", "Please check your 'lib' parameter and try again")) return(list("Bioconductor - lib does not exist")) } } if (file.access(lib,mode=2) < 0) { if (verbose) { print(paste("You do not have write access to",lib, "\nPlease check your permissions or provide", "a different 'lib' parameter")) return(list("Bioconductor - lib is not writeable")) } } biocContribUrl <- sapply(repositories(), contrib.url) biocPkgs <- available.packages(biocContribUrl) if (! cdfname %in% biocPkgs[, "Package"]) { if (verbose) print(paste("Environment",cdfname, "was not found in the Bioconductor", "repository.")) return(list(paste("Bioconductor -",cdfname,"not available"))) } else { install.packages(cdfname, lib=lib, repos=repositories(), dependencies=c("Depends","Imports")) ## no way to know if we succeeded or not, with install.packages ##if (verbose) ## print(paste("Installation of environment", ## cdfname, "was succesful.")) } } else { if (verbose) print(paste("The current operation could not access", "the Bioconductor repository. Please", "check your internet connection, and", "report further problems to", "bioconductor@stat.math.ethz.ch")) return(list("Bioconductor - could not connect")) } return(cdfFromLibPath(cdfname, lib=lib, verbose=verbose)) } cdfFromLibPath <- function(cdfname, lib = NULL, verbose=TRUE) { cdfname <- cleancdfname(cdfname) ## First check to see if package is installed if (verbose) print(paste("Checking to see if package",cdfname, "is already installed")) if (length(find.package(cdfname, lib.loc=lib, quiet=TRUE)) == 0) return(list(paste("Library - package",cdfname,"not installed"))) ## See if package is already loaded if (cdfname %in% .packages()) { if (verbose) print(paste("The package", cdfname, "is already loaded")) } else { if (verbose) print(paste("Attempting to load package", cdfname)) ## Attempt to load the library requested do.call(library, list(cdfname, lib.loc=lib, character.only=TRUE)) ## Check to see if it got loaded if (! cdfname %in% .packages()) { ## package didn't get loaded if (verbose) print(paste("The package", cdfname, "could not be loaded")) return(list(paste("Library - package",cdfname,"is not loadable"))) } } return(get(cdfname, envir=as.environment(paste("package:", cdfname, sep="")))) } affy/R/hlog.R0000644000175200017520000000062514516003533014001 0ustar00biocbuildbiocbuild hlog <- function(x,constant=1){ #constant is where the change occurs if(constant<=0){ warning("constant less than or equal to 0. Returning log(x)\n") return(log(x)) } else{ if(constant==Inf) return(x) else{ aux <- (abs(x)=constant)*(sign(x)*(constant*log(abs(x/constant))+constant)) aux[x==0] <- 0 return(aux) } } } affy/R/justrma.R0000644000175200017520000001172614516003533014541 0ustar00biocbuildbiocbuild## Sept 11, 2003 - justRMA calls just.rma2 ### A user friendly wrapper for just.rma justRMA <- function(..., filenames=character(0), widget=getOption("BioC")$affy$use.widgets, compress=getOption("BioC")$affy$compress.cel, celfile.path=getwd(), sampleNames=NULL, phenoData=NULL, description=NULL, notes="", rm.mask=FALSE, rm.outliers=FALSE, rm.extra=FALSE, hdf5=FALSE, hdf5FilePath=NULL,verbose=FALSE, normalize=TRUE, background=TRUE, bgversion=2, destructive=FALSE, cdfname = NULL){ l <- AllButCelsForReadAffy(..., filenames=filenames, widget=widget, celfile.path=celfile.path, sampleNames=sampleNames, phenoData=phenoData, description=description) ##and now we are ready to read cel files ret<- just.rma(filenames=l$filenames, phenoData=l$phenoData, description=l$description, notes=notes, compress=compress, rm.mask=rm.mask, rm.outliers=rm.outliers, rm.extra=rm.extra, verbose=verbose, normalize=normalize, background=background, bgversion=bgversion, destructive=destructive, cdfname = cdfname) sampleNames(ret) <- l$sampleNames return(ret) } ########################################################################################### # # this function uses a different parsing routine # It was added Jul 7, 2003 by B. M. Bolstad # ########################################################################################### just.rma <- function(..., filenames=character(0), phenoData=new("AnnotatedDataFrame"), description=NULL, notes="", compress=getOption("BioC")$affy$compress.cel, rm.mask=FALSE, rm.outliers=FALSE, rm.extra=FALSE, verbose=FALSE, background=TRUE, normalize=TRUE, bgversion=2, destructive=FALSE, cdfname = NULL) { auxnames <- unlist(list(...)) filenames <- c(filenames, auxnames) checkValidFilenames(filenames) n <- length(filenames) pdata <- pData(phenoData) ##try to read sample names form phenoData. if not there use CEL filenames if(dim(pdata)[1]!=n){#if empty pdata filename are samplenames warning("Incompatible phenoData object. Created a new one.\n") samplenames <- gsub("^/?([^/]*/)*", "", unlist(filenames)) pdata <- data.frame(sample=1:n,row.names=samplenames) phenoData <- new("AnnotatedDataFrame", data=pdata, varMetadata=data.frame( labelDescription="arbitrary numbering", row.names="sample")) } else samplenames <- rownames(pdata) if (is.null(description)) { description <- new("MIAME") description@preprocessing$filenames <- filenames description@preprocessing$affyversion <- as.character(packageVersion("affy")) } ## read the first file to see what we have ##if (verbose) cat(1, "reading",filenames[[1]],"...") ## get information from cdf environment headdetails <- read.celfile.header(filenames[[1]]) if(is.null(cdfname)) cdfname <- headdetails[[1]] scandates <- sapply(seq_len(length(filenames)), function(i) { sdate <- read.celfile.header(filenames[i], info = "full")[["ScanDate"]] if (is.null(sdate) || length(sdate) == 0) NA_character_ else sdate }) protocol <- new("AnnotatedDataFrame", data=data.frame("ScanDate"=scandates, row.names = sampleNames(phenoData), stringsAsFactors=FALSE), dimLabels=c("sampleNames", "sampleColumns")) tmp <- new("AffyBatch", cdfName=cdfname, annotation=cleancdfname(cdfname, addcdf=FALSE)) pmIndex <- pmindex(tmp) probenames <- rep(names(pmIndex), unlist(lapply(pmIndex,length))) pNList <- split(0:(length(probenames) -1), probenames) ## read pm data into matrix probeintensities <- read.probematrix(filenames=filenames, cdfname = cdfname) ##pass matrix of pm values to rma ngenes <- length(geneNames(tmp)) exprs <- .Call("rma_c_complete",probeintensities$pm, pNList, ngenes, normalize, background, bgversion, verbose, PACKAGE="affy") colnames(exprs) <- samplenames se.exprs <- array(NA, dim(exprs), dimnames=list(rownames(exprs), colnames(exprs))) annotation <- annotation(tmp) notes(description) <- notes new("ExpressionSet", phenoData = phenoData, protocolData = protocol, annotation = annotation, experimentData = description, exprs = exprs, se.exprs = se.exprs) } affy/R/loess.normalize.R0000644000175200017520000000276614516003533016204 0ustar00biocbuildbiocbuildloess.normalize <- function(mat,subset=sample(1:(dim(mat)[2]),5000), epsilon=10^-2,maxit=1,log.it=TRUE,verbose=TRUE,span=2/3, family.loess="symmetric") { .Deprecated("normalize.loess", "affy") J <- dim(mat)[2] II <- dim(mat)[1] newData <- mat if(log.it){ mat <- log2(mat) newData <- log2(newData) } change <- epsilon +1 fs <- matrix(0,II,J)##contains what we substract iter <- 0 w <- c(0,rep(1,length(subset)),0) ##this way we give 0 weight to the ##extremes added so that we can interpolate while(iter < maxit){ iter <- iter+1 means <- matrix(0,II,J) ##contains temp of what we substract for(j in 1:(J-1)){ for(k in (j+1):J){ y <- newData[,j]-newData[,k] x <-(newData[,j]+newData[,k])/2 index <- c(order(x)[1],subset,order(-x)[1]) ##put endpoints in so we can interpolate xx <- x[index] yy <- y[index] aux <-loess(yy~xx,span=span,degree=1,weights=w,family=family.loess) aux <- predict(aux,data.frame(xx=x))/J means[,j] <- means[,j] + aux means[,k] <- means[,k] - aux if(verbose) cat("Done with",j,"vs",k," in iteration ",iter,"\n") } } fs <- fs+means newData <- mat-fs change <- max(colMeans((means[subset,])^2)) if(verbose) cat(iter,change,"\n") oldfs <- fs } if(change>epsilon & maxit>1) warning(paste("No convergence after",maxit,"iterations.\n")) if(log.it) return(2^newData) else return(newData) } affy/R/maffy.R0000644000175200017520000002500714516003533014153 0ustar00biocbuildbiocbuild##******************************************************************************************* #********** maffy.normalize ***** maffy.normalize <- function(data,subset,verbose=FALSE,span=0.25,family="symmetric",log.it=TRUE){ k <- dim(data)[2] ### Number of chips #### Create the transformation matrix t1 <- 1/sqrt(k) t2 <- (k-2-t1)/(k-1) t3 <- -(1+t1)/(k-1) transmat <- matrix(t3,k,k) for(i in 1:k){ transmat[1,i]<-t1 transmat[i,1]<-t1 } for(i in 2:k) transmat[i,i]<-t2 #### Find normalizing curve if(verbose) cat("Fitting normalizing curve\n") n<- length(subset) data.subset <- data[subset,] data.subset <- log(data.subset)%*%t(transmat) index <- order(data.subset[,1]) data.subset <- data.subset[index,] if( k>2) curve <- multiloess(data.subset[,2:k]~data.subset[,1],span=span,family=family,surface="direct") else curve <- loess(data.subset[,2:k]~data.subset[,1],span=span,family=family,surface="direct") ### Transform the normalizing curve before and after normalization scaled <- cbind(data.subset[,1],matrix(0,n,k-1)) %*%(transmat) unscaled <- cbind(data.subset[,1],curve$fitted) %*%(transmat) w <-c(0,rep(1,n,n),0) data.scaled <- NULL ### Normalize each array for(i in 1:k){ if(verbose) cat("Normalizing chip ",i,"\n") if(log.it){ mini <- log(min(data[,i])) maxi <- log(max(data[,i])) } else{ mini <- min(data[,i]) maxi <- max(data[,i]) } curve <- loess(c(mini,scaled[,i],maxi)~c(mini,unscaled[,i],maxi),weights=w,span=span) if(log.it) temp <- exp(predict(curve,log(data[,i]))) else temp <- predict(curve,data[,i]) data.scaled <- cbind(data.scaled,temp) } data.scaled } ##******************************************************************************************* #********** Select A subset with small rank-range over arrays ***** maffy.subset <- function(data,subset.size=5000,maxit=100,subset.delta=max(round(subset.size/100),25),verbose=FALSE){ k <- dim(data)[2] ### Number of chips n <- dim(data)[1] ## Size of starting subset, i.e. all rows if(verbose) cat("Data size",n,"x",k,"Desired subset size",subset.size,"+-",subset.delta,"\n") means <- data%*%(rep(1,k,k)/k) index0 <- order(means) data.sorted <- data[index0,] ## Init set <- rep(TRUE,n,n) ## Set-indicator index.set <- 1:n ## Indexes for subset nprev <- n+1 iter <- 1 part.of.n <- 1 ## loop while(nprev>n & n>(subset.size+subset.delta) & iter 0 parametric <- match(nmx, nmx[parametric], 0) > 0 if(!match(degree, 0:2, 0)) stop("degree must be 0, 1 or 2") iterations <- if(family=="gaussian") 1 else control$iterations if(!missing(enp.target)) if(!missing(span)) warning("both span and enp.target specified: span will be used") else { # White book p.321 tau <- switch(degree+1, 1, D+1, (D+1)*(D+2)/2) - sum(drop.square) span <- 1.2 * tau/enp.target } fit <- simplemultiLoess(y, x, w, span, degree, normalize, control$statistics, control$surface, control$cell, iterations, control$trace.hat) fit$call <- match.call() fit$terms <- mt fit$xnames <- nmx fit$x <- x fit$y <- y fit$weights <- w if(model) fit$model <- mf fit } ##******************************************************************************************* simplemultiLoess <- function(y, x, weights, span = 0.75, degree = 2, normalize = TRUE, statistics = "approximate", surface = "interpolate", cell = 0.2, iterations = 1, trace.hat = "exact") { ## Extra init parametric <- FALSE drop.square <- FALSE M <- NCOL(y) A <- rep(1,M,M) D <- NCOL(x) N <- NROW(x) fitted.all <- matrix(1,N,M) fitted.residuals <- matrix(1,N,M) pseudo.resid.all <- matrix(1,N,M) if(!N || !D) stop("invalid `x'") if(!length(y)) stop("invalid `y'") x <- as.matrix(x) max.kd <- max(N, 200) robust <- rep(1, N) divisor<- rep(1, D) if(normalize && D > 1) { trim <- ceiling(0.1 * N) divisor <- sqrt(apply(apply(x, 2, sort)[seq(trim+1, N-trim), , drop = FALSE], 2, var)) x <- x/rep(divisor, rep(N, D)) } sum.drop.sqr <- sum(drop.square) sum.parametric <- sum(parametric) nonparametric <- sum(!parametric) order.parametric <- order(parametric) x <- x[, order.parametric] order.drop.sqr <- (2 - drop.square)[order.parametric] if(degree==1 && sum.drop.sqr) stop("Specified the square of a factor predictor to be dropped when degree = 1") if(D == 1 && sum.drop.sqr) stop("Specified the square of a predictor to be dropped with only one numeric predictor") if(sum.parametric == D) stop("Specified parametric for all predictors") if(iterations) for(j in 1:iterations) { robust <- weights * robust if(j > 1) statistics <- "none" if(surface == "interpolate" && statistics == "approximate") statistics <- if(trace.hat == "approximate") "2.approx" else if(trace.hat == "exact") "1.approx" surf.stat <- paste(surface, statistics, sep="/") for(k in 1:M) { z <- .C(stats:::C_loess_raw, as.double(y[,k]), as.double(x), as.double(weights), as.double(robust), as.integer(D), as.integer(N), as.double(span), as.integer(degree), as.integer(nonparametric), as.integer(order.drop.sqr), as.integer(sum.drop.sqr), as.double(span*cell), as.character(surf.stat), fitted.values = double(N), parameter = integer(7), a = integer(max.kd), xi = double(max.kd), vert = double(2*D), vval = double((D+1)*max.kd), diagonal = double(N), trL = double(1), delta1 = double(1), delta2 = double(1), as.integer(surf.stat == "interpolate/exact")) fitted.all[,k] <- z$fitted.values } if(j==1) { trace.hat.out <- z$trL one.delta <- z$delta1 two.delta <- z$delta2 } residuals.all <- (y-fitted.all) fitted.residuals <- sqrt((residuals.all^2)%*%A) if(j < iterations) robust <- .Fortran(stats:::C_lowesw, as.double(fitted.residuals), as.integer(N), robust = double(N), integer(N))$robust } if(surface == "interpolate") { pars <- z$parameter names(pars) <- c("d", "n", "vc", "nc", "nv", "liv", "lv") enough <- (D + 1) * pars["nv"] fit.kd <- list(parameter=pars, a=z$a[1:pars[4]], xi=z$xi[1:pars[4]], vert=z$vert, vval=z$vval[1:enough]) } if(iterations > 1) { for(k in 1:M) { pseudovalues <- .Fortran(stats:::C_lowesp, as.integer(N), as.double(y[,k]), as.double(fitted.all[,k]), as.double(weights), as.double(robust), integer(N), pseudovalues = double(N))$pseudovalues zz <- .C(stats:::C_loess_raw, as.double(pseudovalues), as.double(x), as.double(weights), as.double(weights), as.integer(D), as.integer(N), as.double(span), as.integer(degree), as.integer(nonparametric), as.integer(order.drop.sqr), as.integer(sum.drop.sqr), as.integer(span*cell), as.character(surf.stat), temp = double(N), parameter = integer(7), a = integer(max.kd), xi = double(max.kd), vert = double(2*D), vval = double((D+1)*max.kd), diagonal = double(N), trL = double(1), delta1 = double(1), delta2 = double(1), as.integer(0)) pseudo.resid.all[,k] <- pseudovalues-zz$temp } pseudo.resid <- sqrt((pseudo.resid.all^2)%*%A) } sum.squares <- if(iterations <= 1) sum(weights * fitted.residuals^2) else sum(weights * pseudo.resid^2) enp <- one.delta + 2*trace.hat.out - N s <- sqrt(sum.squares/one.delta) pars <- list(robust=robust, span=span, degree=degree, normalize=normalize, parametric=parametric, drop.square=drop.square, surface=surface, cell=cell, family= if(iterations <= 1) "gaussian" else "symmetric", iterations=iterations) fit <- list(n=N, fitted=fitted.all, residuals=residuals.all, enp=enp, s=s, one.delta=one.delta, two.delta=two.delta, trace.hat=trace.hat.out, divisor=divisor) fit$pars <- pars if(surface == "interpolate") fit$kd <- fit.kd class(fit) <- "loess" fit } ##******************************************************************************************* affy/R/mas5.R0000644000175200017520000001305214516003533013713 0ustar00biocbuildbiocbuildmas5 <- function(object,normalize=TRUE,sc = 500, analysis = "absolute",...){ res <- expresso(object,bgcorrect.method="mas",pmcorrect.method="mas",normalize=FALSE,summary.method="mas",...) if(normalize) res <- affy.scalevalue.exprSet(res,sc=sc,analysis=analysis) return(res) } mas5calls.ProbeSet <- function(object, tau=0.015, alpha1=0.04, alpha2=0.06, ignore.saturated=TRUE){ if(alpha1 < 0) {stop("alpha1 must be > 0 "); } if(alpha1 > alpha2) {stop("alpha2 must be > alpha1 "); } if(alpha2 > 1) {stop("alpha2 must be <1 "); } ## Saturation: ## shouldn't be a problem with new scanners ##or those that have had an engineer visit if(ignore.saturated) { sat <- 46000; } else { sat <- -1; } pms <- pm(object) mms <- mm(object) pns <- rep(object@id,nrow(pms)) unique.pns <- unique(pns) pvals<-sapply(1:length(pms[1,]),function(x) { .C("DetectionPValue",as.double(pms[,x]),as.double(mms[,x]),as.character(pns),as.integer(length(mms[,x])), as.double(tau),as.double(sat),dpval=double(length(unique.pns)),length(unique.pns), PACKAGE="affy")$dpval; }); calls <- sapply(pvals,function(y) { if(y < alpha1) { return("P") } else { if(y < alpha2) { return("M") } else { return("A") }}}); return(list(call=calls,pval=pvals)) } mas5calls.AffyBatch <- function(object, ids=NULL, verbose=TRUE, tau=0.015, alpha1=0.04, alpha2=0.06, ignore.saturated=TRUE) { if(alpha1 < 0) {stop("alpha1 must be > 0 "); } if(alpha1 > alpha2) {stop("alpha2 must be > alpha1 "); } if(alpha2 > 1) {stop("alpha2 must be <1 "); } if(verbose) cat("Getting probe level data...\n"); pms <-as.matrix(pm(object)); mms <-as.matrix(mm(object)); # Saturation: # shouldn't be a problem with new scanners ##or those that have had an engineer visit if(ignore.saturated) { sat <- 46000; } else { sat <- -1; } pns <- probeNames(object); o <- order(pns) pns <- pns[o] pms <- pms[o,,drop=FALSE] mms <- mms[o,,drop=FALSE] unique.pns <- sort(unique(pns)); if(verbose) cat("Computing p-values\n"); p<-sapply(1:length(pms[1,]),function(x) { .C("DetectionPValue",as.double(pms[,x]),as.double(mms[,x]),as.character(pns),as.integer(length(mms[,x])), as.double(tau),as.double(sat),dpval=double(length(unique.pns)),length(unique.pns), PACKAGE="affy")$dpval; }); rownames(p) <- unique.pns; colnames(p) <- sampleNames(object) if(verbose) cat("Making P/M/A Calls\n"); calls <- sapply(p,function(y) { if(y < alpha1) { return("P") } else { if(y < alpha2) { return("M") } else { return("A") }}}); calls <- matrix(calls,nrow=nrow(p),ncol=ncol(p)); colnames(calls) <- sampleNames(object) rownames(calls) <- rownames(p) if(!is.null(ids)){ calls <- calls[ids,,drop=FALSE] p <- p[ids,,drop=FALSE] } eset <- new("ExpressionSet", phenoData=phenoData(object), ## featureData picked up from object experimentData=experimentData(object), annotation=annotation(object), protocolData=protocolData(object), exprs=calls, se.exprs=p ) return(eset) } mas5.detection <- function(mat, tau=0.015, alpha1=0.04, alpha2=0.06, exact.pvals=FALSE, cont.correct=FALSE) { ## CONSTANTS saturation.point <- 46000 # not a user parameter mat.r <- (mat[,1]-mat[,2])/(mat[,1]+mat[,2]) ## SANITY CHECKING if ( !is.matrix(mat) || length(dim(mat))!=2 || dim(mat)[2]!=2 || dim(mat)[1] < 1 || !is.numeric(mat) ) stop("Invalid mat matrix.") if ( !is.numeric(tau) ) stop("Invalid tau.") if ( !is.numeric(alpha1) || !is.numeric(alpha2) || alpha1 <= 0 || alpha1 >= alpha2 || alpha2 >= 0.5 ) stop("Invalid alpha1 or alpha2.") if ( !is.logical(exact.pvals) ) stop("Invalid exact.pvals.") if ( !is.logical(cont.correct) ) stop("Invalid cont.correct.") ## DEALING WITH SATURATION; COMPUTING THE P-VALUE ## According to the Bioinformatics paper: ## * If all MM's are saturated, then call present ## * Otherwise discard pairs with a saturated MM ## According to the Affymetrix whitepaper: ## * If all probe-pairs are saturated, then call present with pval=0 ## * If an MM is saturated, then we discard the pair ## * If a PM and MM are within tau of each other, we discard the pair ## So we're going with: ## * If all MM's are saturated, set pval=0 and don't use Wilcoxon ## * Discard probe-pairs when MM is saturated or the PM,MM are within tau ## of each other ## * Compute the p-value using Wilcoxon's signed rank test on the retained ## probe-pairs is.mm.saturated <- function(probe.pair, saturation.point) probe.pair[2] >= saturation.point is.retained <- function(probe.pair, saturation.point, tau) !(is.mm.saturated(probe.pair,saturation.point) || abs(diff(probe.pair)) <= tau) if ( all(apply(mat,1,is.mm.saturated,saturation.point)) ) pval <- 0 else { retained <- apply(mat, 1, is.retained, saturation.point, tau) pval <- wilcox.test(mat.r[retained], alternative="greater", mu=tau, paired=FALSE, exact=exact.pvals, correct=cont.correct, conf.int=FALSE)$p.value } ## DETECTION CALL if ( pval < 0 || pval > 1 ) warning("Computed an unusual p-value outside the range [0,1].") if ( pval < alpha1 ) call <- "P" else if ( pval < alpha2 ) call <- "M" else call <- "A" ## DONE return(list(pval=pval, call=call)) } affy/R/merge.AffyBatch.R0000644000175200017520000000264014516003533015774 0ustar00biocbuildbiocbuildmerge.AffyBatch <- function(x, y, annotation=paste(annotation(x), annotation(y)), description=NULL, notes=character(0), ...) { adim <- dim(intensity(x))[1] if ((nrow(x) != nrow(y)) || (ncol(x) != ncol(y))) stop("cannot merge chips of different sizes !") if (cdfName(x) != cdfName(y)) warning("cdfName mismatch (using the cdfName of x)!") if (is.null(description)){ description <- new("MIAME") description@title <- "Created from merging two AffyBatches. No description was supplied. The description of the two original AffyBatches was not kept." } lx <- length(x) ly <- length(y) phenodata <- phenoData(x) pData(phenodata) <- rbind(pData(x),pData(y)) protocoldata <- protocolData(x) pData(protocoldata) <- rbind(pData(protocolData(x)),pData(protocolData(y))) notes(description) <- if (length(notes)==0) list(paste("Merge from two AffyBatches with notes: 1)", notes(experimentData(x)), ", and 2)",notes(experimentData(y)))) else notes return(new("AffyBatch", exprs=cbind(intensity(x),intensity(y)), phenoData=phenodata, experimentData=description, ##need to write a merge for MIAME cdfName=cdfName(x), nrow=nrow(x), ncol=ncol(x), annotation=x@annotation, protocolData=protocoldata )) } affy/R/mva.pairs.R0000644000175200017520000003440614516003533014754 0ustar00biocbuildbiocbuild### ### ### Code for M and MvA plots ### ### Mar 6, 2004 - added the generic Mbox. It performs ### the equivalent of Mbox in affyPLM ### added a generic MAplot. Similar ### functionality is implemented in ### affyPLM ### a function ma.plot now does the actual plotting ### for mva.pairs ### ### Aug 23, 2004 - change the placement location of statistics in ### ma.plot ### Nov 30, 2005 - fix double logging when pairs=TRUE in MAplot ### Feb 15, 2006 - fixed passing of cex variable into mva.pairs ### Feb 24, 2006 - add smoothScatter option to ma.plot ### Apr 11, 2006 - fix problem with smoothScatter option. ### Jun 22, 2006 - Fix problem with where statistics appear when xlim is set. Add plotmethod="add". move pch to MAplot. Fix it so cex is passed down to plot(). Make adding the loess line optional. Make lwd,lty,col settable for the loess line ### Jul 21, 2006 - allow MAplot to have character string sampleName arguments for which,subset,ref. But then removed subset ### Jul 23, 2006 - added groups to MAplot for AffyBatch ### Aug 4, 2006 - fix small bug in how plots are titled. ### Oct 11, 2006 - change some apply(x,1,median) to rowMedians(x) ### ma.plot <- function(A,M,subset=sample(1:length(M),min(c(10000, length(M)))),show.statistics=TRUE,span=2/3,family.loess="gaussian",cex=2,plot.method=c("normal","smoothScatter","add"),add.loess=TRUE,lwd=1,lty=1,loess.col="red",...){ plot.method <- match.arg(plot.method) fn.call <- list(...) sigma <- IQR(M) mean <- median(M) if (!is.element("ylim",names(fn.call))){ yloc <- max(M) } else { yloc <- max(fn.call$ylim) } if (!is.element("xlim",names(fn.call))){ xloc <- max(A) } else { xloc <- max(fn.call$xlim) } if(plot.method == "smoothScatter"){ plotmethod <- "smoothScatter" } else if (plot.method == "add"){ plotmethod <- "add" } else { plotmethod <- "normal" } aux <- loess(M[subset]~A[subset],degree=1,span=span,family=family.loess)$fitted if (plotmethod == "smoothScatter"){ smoothScatter(A,M,...) } else if (plotmethod == "add"){ points(A,M,cex=cex,...) } else { plot(A,M,cex=cex,...) } if (add.loess){ o <- order(A[subset]) A <- A[subset][o] M <- aux[o] o <-which(!duplicated(A)) lines(approx(A[o],M[o]),col=loess.col,lwd=lwd,lty=lty) } abline(0,0,col="blue") # write IQR and Median on to plot if (show.statistics){ txt <- format(sigma,digits=3) txt2 <- format(mean,digits=3) text(xloc ,yloc,paste(paste("Median:",txt2),paste("IQR:",txt),sep="\n"),cex=cex,adj=c(1,1)) } } mva.pairs <- function(x,labels=colnames(x),log.it=TRUE,span=2/3,family.loess="gaussian", digits=3,line.col=2,main="MVA plot", cex = 2, ...){ if(log.it) x <-log2(x) J <- dim(x)[2] frame() old.par <- par(no.readonly = TRUE) on.exit(par(old.par)) par(mfrow=c(J,J),mgp=c(0,.2,0),mar=c(1,1,1,1),oma=c(1,1.4,2,1)) for(j in 1:(J-1)){ par(mfg=c(j,j)) plot(1,1,type="n",xaxt="n",yaxt="n",xlab="",ylab="") text(1,1,labels[j],cex=cex) for(k in (j+1):J){ par(mfg=c(j,k)) yy <- x[,j]-x[,k] xx <-(x[,j]+x[,k])/2 sigma <- IQR(yy) mean <- median(yy) ma.plot(xx,yy,tck=0,show.statistics=FALSE,pch=".",xlab="",ylab="",tck=0,span=span,...) par(mfg=c(k,j)) #sigma <- IQR(yy) txt <- format(sigma,digits=digits) txt2 <- format(mean,digits=digits) plot(c(0,1),c(0,1),type="n",ylab="",xlab="",xaxt="n",yaxt="n") text(0.5,0.5,paste(paste("Median:",txt2),paste("IQR:",txt),sep="\n"),cex=cex) } } par(mfg=c(J,J));plot(1,1,type="n",xaxt="n",yaxt="n",xlab="",ylab=""); text(1,1,labels[J],cex=cex) mtext("A",1,outer=TRUE,cex=1.5) mtext("M",2,outer=TRUE,cex=1.5,las=1) mtext(main,3,outer=TRUE,cex=1.5) invisible() } setGeneric("Mbox",function(object,...) standardGeneric("Mbox")) setMethod("Mbox",signature("AffyBatch"), function(object,log=TRUE,type=c("both","pm","mm"),...){ type <- match.arg(type) if (type == "both"){ pms <- unlist(indexProbes(object, "both")) } else if (type == "pm"){ pms <- unlist(pmindex(object)) } else if (type == "mm"){ mms <- unlist(mmindex(object)) } if(log){ x <- log2(intensity(object)[pms, ]) } else { x <- intensity(object)[pms, ] } medianchip <- rowMedians(x) M <- sweep(x,1,medianchip,FUN='-') boxplot(data.frame(M),...) }) setGeneric("MAplot",function(object,...) standardGeneric("MAplot")) setMethod("MAplot",signature("AffyBatch"), function(object,log=TRUE,type=c("both","pm","mm"),groups=NULL,ref=NULL,which=NULL,pairs=FALSE,pch=".",ref.fn=c("median","mean"),ref.title="vs pseudo-median reference chip",...){ type <- match.arg(type) if (type == "both"){ pms <- unlist(indexProbes(object, "both")) } else if (type == "pm"){ pms <- unlist(pmindex(object)) } else if (type == "mm"){ pms <- unlist(mmindex(object)) } if(log){ x <- log2(intensity(object)[pms, ]) } else { x <- intensity(object)[pms, ] } if (is.null(groups)){ if (is.character(ref)){ ref.indices <- match(ref,sampleNames(object)) if (all(is.na(ref.indices))){ stop("No known sampleNames in ref") } if (any(is.na(ref.indices))){ warning(paste("Omitting the following from ref:",ref[is.na(ref.indices)], "because they can not be found.")) } ref <- ref.indices[!is.na(ref.indices)] } if (is.character(subset)){ subset.indices <- match(subset,sampleNames(object)) if (all(is.na(subset.indices))){ stop("No known sampleNames in subset") } if (any(is.na(subset.indices))){ warning(paste("Omitting the following from subset:",subset[is.na(subset.indices)], "because they can not be found.")) } subset <- subset.indices[!is.na(subset.indices)] } if (is.character(which)){ which.indices <- match(which,sampleNames(object)) if (all(is.na(which.indices))){ stop("No known sampleNames in which") } if (any(is.na(which.indices))){ warning(paste("Omitting the following from which:",which[is.na(which.indices)], "because they can not be found.")) } which <- which.indices[!is.na(which.indices)] } if (is.null(which)){ which <- 1:dim(exprs(object))[2] } ref.fn <- match.arg(ref.fn) if(!pairs){ if (is.null(ref)){ medianchip <- rowMedians(x) ###apply(x, 1, median) } else if (length(ref) > 1){ if (ref.fn == "median"){ medianchip <- rowMedians(x[,ref]) } else { medianchip <- rowMeans(x[,ref]) } } else { medianchip <- x[,ref] } M <- sweep(x,1,medianchip,FUN='-') A <- 1/2*sweep(x,1,medianchip,FUN='+') if (is.null(ref)){ for (i in which){ title <- paste(sampleNames(object)[i],"vs pseudo-median reference chip") ma.plot(A[,i],M[,i],main=title,xlab="A",ylab="M",pch=pch,...) } } else { for (i in which){ if (length(ref) == 1){ if (i != ref){ ##changed which to i title <- paste(sampleNames(object)[i],"vs",sampleNames(object)[ref]) ma.plot(A[,i],M[,i],main=title,xlab="A",ylab="M",pch=pch,...) } } else { title <- paste(sampleNames(object)[i],"vs",ref.title) ma.plot(A[,i],M[,i],main=title,xlab="A",ylab="M",pch=pch,...) } } } } else { if(!is.null(ref)) stop("Can't use pairs with non-null 'ref'") if(is.null(which)) which <- 1:ncol(x) mva.pairs(x[,which],log.it=FALSE,...) } } else { ## group labels have been given ## check that group variable is of same length as number of samples if (dim(x)[2] != length(groups)){ stop("'groups' is of wrong length.") } ### group labels variable can be integer, character or factor variable. ### need to check that if any names supplied ### for ref or which can be found in group.labels if (!is.null(which)){ if (is.numeric(groups)){ if (!is.numeric(which)){ stop("'which' labels must also be found in 'groups'") } else { if (!all(is.element(which,groups))){ stop("'which' labels must also be found in 'groups'") } } } else if (is.factor(groups)){ if (!is.character(which)){ stop("'which' should be character vector") } else { if (!all(is.element(which,as.character(groups)))){ stop("'which' labels must also be found in 'groups'") } } } else if (is.character(groups)){ if (!is.character(which)){ stop("'which' should be character vector") } else { if (!all(is.element(which,groups))){ stop("'which' labels must also be found in 'groups'") } } } } if (!is.null(ref)){ if (is.numeric(groups)){ if (!is.numeric(ref)){ stop("'ref' labels must also be found in 'groups'") } else { if (!all(is.element(ref,groups))){ stop("'ref' labels must also be found in 'groups'") } } } else if (is.factor(groups)){ if (!is.character(ref)){ stop("'ref' should be character vector") } else { if (!all(is.element(ref,as.character(groups)))){ stop("'ref' labels must also be found in 'groups'") } } } else if (is.character(groups)){ if (!is.character(ref)){ stop("'ref' should be character vector") } else { if (!all(is.element(ref,groups))){ stop("'ref' labels must also be found in 'groups'") } } } } ref.fn <- match.arg(ref.fn) groups.list <- split(1:dim(x)[2], as.factor(groups)) grouped.data <- matrix(0,nrow(x),length(groups.list)) colnames(grouped.data) <- names(groups.list) which.col <- 1 for (group in groups.list){ grouped.data[,which.col] <- rowMeans(x[,group,drop=FALSE]) which.col <- which.col + 1 } if (!pairs){ if (is.null(which)){ which <- names(groups.list) } if (is.null(ref)){ if (ref.fn == "median"){ medianchip <- rowMedians(grouped.data) ####apply(grouped.data, 1, median) } else { medianchip <- rowMeans(grouped.data) } } else if (length(ref) == 1){ ref.name <- ref ref <- match(ref,names(groups.list)) medianchip <- grouped.data[,ref] } else { ref <- match(ref,names(groups.list)) if (ref.fn == "median"){ medianchip <- rowMedians(grouped.data[,ref]) } else { medianchip <- rowMeans(grouped.data[,ref]) } } M <- sweep(grouped.data,1,medianchip,FUN='-') A <- 1/2*sweep(grouped.data,1,medianchip,FUN='+') if (is.null(ref)){ for (i in which){ title <- paste(i,ref.title) ma.plot(A[,i],M[,i],main=title,xlab="A",ylab="M",pch=pch,...) } } else { for (i in which){ if (length(ref) == 1){ if (i != ref.name){ title <- paste(i,"vs",ref.name) ma.plot(A[,i],M[,i],main=title,xlab="A",ylab="M",pch=pch,...) } } else { title <- paste(i,ref.title) ma.plot(A[,i],M[,i],main=title,xlab="A",ylab="M",pch=pch,...) } } } } else { if (!is.null(ref)) stop("Can't use pairs with non-null 'ref'") if (is.null(which)){ which <- names(groups.list) } mva.pairs(grouped.data[,which],log.it=FALSE,...) } } }) affy/R/normalize.constant.R0000644000175200017520000000165314516003533016702 0ustar00biocbuildbiocbuildnormalize.AffyBatch.constant <- function(abatch, refindex=1, FUN=mean, na.rm=TRUE) { n <- length( abatch ) if (! (refindex %in% 1:n)) stop("invalid reference index for normalization") refconstant <- FUN(intensity(abatch)[,refindex], na.rm=na.rm) #set.na.spotsd(abatch) normhisto <- vector("list", length=n) for (i in (1:n)[-refindex]) { m <- normalize.constant(intensity(abatch)[,i], refconstant, FUN=FUN, na.rm=na.rm) myhistory <- list(name="normalized by constant", constant=attr(m,"constant")) attr(m,"constant") <- NULL intensity(abatch)[, i] <- m normhisto[[i]] <- myhistory } attr(abatch, "normalization") <- normhisto return(abatch) } normalize.constant <- function(x, refconstant, FUN=mean, na.rm=TRUE) { thisconstant <- FUN(x, na.rm=na.rm) r <- x / thisconstant * refconstant attr(r,"constant") <- refconstant / thisconstant return(r) } affy/R/normalize.contrasts.R0000644000175200017520000000217114516003533017065 0ustar00biocbuildbiocbuildnormalize.AffyBatch.contrasts <- function(abatch,span=2/3,choose.subset=TRUE,subset.size=5000,verbose=TRUE,family="symmetric",type=c("together","pmonly","mmonly","separate")) { type <- match.arg(type) if (type == "pmonly"){ Index <- unlist(pmindex(abatch)) } else if (type == "mmonly"){ Index <- unlist(mmindex(abatch)) } else if (type == "together"){ Index <- unlist(indexProbes(abatch,"both")) } else if (type == "separate"){ abatch <- normalize.AffyBatch.contrasts(abatch,span=span,choose.subset=choose.subset,subset.size=subset.size,verbose=verbose,family=family,type="pmonly") Index <- unlist(mmindex(abatch)) } ##we need default argumetns becuase they are used in this transitional file alldata <- intensity(abatch)[Index,] if(choose.subset) subset1 <- maffy.subset(alldata,verbose=verbose,subset.size=subset.size)$subset else subset1 <- sample(1:dim(alldata)[1],subset.size) aux <- maffy.normalize(alldata,subset=subset1,verbose=verbose,span=span,family=family) intensity(abatch)[Index,] <- aux ##attr(abatch, "normalization") <- normhisto return(abatch) } affy/R/normalize.invariantset.R0000644000175200017520000001227014516003533017555 0ustar00biocbuildbiocbuildnormalize.AffyBatch.invariantset <- function(abatch, prd.td=c(0.003,0.007), verbose=FALSE,baseline.type=c("mean","median","pseudo-mean","pseudo-median"),type=c("separate","pmonly","mmonly","together")) { do.normalize.Affybatch.invariantset <- function(abatch, pms, prd.td, baseline.type){ nc <- length(abatch) # number of CEL files if (baseline.type == "mean"){ # take as a reference the array having the median overall intensity m <- vector("numeric", length=nc) for (i in 1:nc) m[i] <- mean(intensity(abatch)[pms, i]) refindex <- match(trunc(median(rank(m))), rank(m)) rm(m) baseline.chip <- c(intensity(abatch)[pms, refindex]) if (verbose) cat("Data from", sampleNames(abatch)[refindex], "used as baseline.\n") } else if (baseline.type == "median"){ # take as a reference the array having the median median intensity m <- vector("numeric", length=nc) for (i in 1:nc) m[i] <- median(intensity(abatch)[pms, i]) refindex <- match(trunc(median(rank(m))), rank(m)) rm(m) baseline.chip <- c(intensity(abatch)[pms, refindex]) if (verbose) cat("Data from", sampleNames(abatch)[refindex], "used as baseline.\n") } else if (baseline.type == "pseudo-mean"){ # construct a psuedo chip to serve as the baseline by taking probewise means refindex <- 0 baseline.chip <- rowMeans(intensity(abatch)[pms,]) } else if (baseline.type == "pseudo-median"){ # construct a pseudo chip to serve as the baseline by taking probewise medians refindex <- 0 baseline.chip <- rowMedians(intensity(abatch)[pms,]) } ##set.na.spotsd(cel.container) normhisto <- vector("list", length=nc) # normhisto[[refindex]] <- list(name="reference for the invariant set") ## loop over the CEL files and normalize them for (i in (1:nc)) { if (i != refindex){ if (verbose) cat("normalizing array", sampleNames(abatch)[i], "...") ##temporary tmp <- normalize.invariantset(c(intensity(abatch)[pms, i]), c(baseline.chip), prd.td) #i.set <- which(i.pm)[tmp$i.set] tmp <- as.numeric(approx(tmp$n.curve$y, tmp$n.curve$x, xout=intensity(abatch)[pms, i], rule=2)$y) attr(tmp,"invariant.set") <- NULL intensity(abatch)[pms, i] <- tmp ## storing information about what has been done #normhisto[[i]] <- list(name="normalized by invariant set", # invariantset=i.set) if (verbose) cat("done.\n") } } attr(abatch, "normalization") <- normhisto return(abatch) } type <- match.arg(type) baseline.type <- match.arg(baseline.type) if (type == "pmonly"){ pms <- unlist(pmindex(abatch)) do.normalize.Affybatch.invariantset(abatch, pms, prd.td, baseline.type) } else if (type == "mmonly"){ pms <- unlist(mmindex(abatch)) do.normalize.Affybatch.invariantset(abatch, pms, prd.td, baseline.type) } else if (type == "together"){ pms <- unlist(indexProbes(abatch,"both")) do.normalize.Affybatch.invariantset(abatch, pms, prd.td, baseline.type) } else if (type == "separate"){ pms <- unlist(pmindex(abatch)) abatch <- do.normalize.Affybatch.invariantset(abatch, pms, prd.td, baseline.type) pms <- unlist(mmindex(abatch)) do.normalize.Affybatch.invariantset(abatch, pms, prd.td, baseline.type) } } ## The 'common-to-all' part of the algorithm. Operates on two vectors of numeric data ## normalize.invariantset <- function(data, ref, prd.td=c(0.003,0.007)) { np <- length(data) r.ref <- rank(ref) r.array <- rank(data) ## init prd.td.adj <- prd.td*10 # adjusted threshold things i.set <- rep(TRUE, np) # index all the PM probes as being in the invariant set ns <- sum(i.set) # number of probes in the invariant set ns.old <- ns+50+1 # number of probes previously in the invariant set ## iterate while the number of genes in the invariant set (ns) still varies... while ( (ns.old-ns) > 50 ) { air <- (r.ref[i.set] + r.array[i.set]) / (2*ns) # average intensity rank for the probe intensities prd <- abs(r.ref[i.set] - r.array[i.set]) / ns threshold <- (prd.td.adj[2]-prd.td[1]) * air + prd.td.adj[1] i.set[i.set] <- (prd < threshold) ns.old <- ns ns <- sum(i.set) if (prd.td.adj[1] > prd.td[1]) prd.td.adj <- prd.td.adj * 0.9 # update the adjusted threshold parameters } ## the index i.set corresponds to the 'invariant genes' n.curve <- smooth.spline(ref[i.set], data[i.set]) ## n.curve$x contains smoothed reference intensities ## n.curve$y contains smoothed i-th array intensities ##data <- as.numeric(approx(n.curve$y, n.curve$x, xout=data)$y) ##attr(data,"invariant.set") <- i.set ##return(data) return(list(n.curve=n.curve, i.set=i.set)) } affy/R/normalize.loess.R0000644000175200017520000000506614516003533016200 0ustar00biocbuildbiocbuildnormalize.AffyBatch.loess <- function(abatch,type=c("together","pmonly","mmonly","separate"),...) { type <- match.arg(type) if (type == "separate"){ Index <- unlist(indexProbes(abatch,"pm")) intensity(abatch)[Index,] <- normalize.loess(intensity(abatch)[Index,], ...) Index <- unlist(indexProbes(abatch,"mm")) intensity(abatch)[Index,] <- normalize.loess(intensity(abatch)[Index,], ...) } else if (type=="together"){ Index <- unlist(indexProbes(abatch,"both")) intensity(abatch)[Index,] <- normalize.loess(intensity(abatch)[Index,], ...) } else if (type=="pmonly"){ Index <- unlist(indexProbes(abatch,"pm")) intensity(abatch)[Index,] <- normalize.loess(intensity(abatch)[Index,], ...) } else if (type=="mmonly"){ Index <- unlist(indexProbes(abatch,"mm")) intensity(abatch)[Index,] <- normalize.loess(intensity(abatch)[Index,], ...) } ##set.na.spotsd(listcel) # set 'sd' to nothing (meaningless after normalization) ##cat(cols,rows) ##need to use MIAME ##for (i in 1:abatch@nexp) { ## history(abatch)[[i]] <- list(name="normalized by loess") ##} return(abatch) } normalize.loess <- function(mat, subset=sample(1:(dim(mat)[1]), min(c(5000, nrow(mat)))), epsilon=10^-2, maxit=1, log.it=TRUE, verbose=TRUE, span=2/3, family.loess="symmetric"){ J <- dim(mat)[2] II <- dim(mat)[1] if(log.it){ mat <- log2(mat) } change <- epsilon +1 iter <- 0 w <- c(0, rep(1,length(subset)), 0) ##this way we give 0 weight to the ##extremes added so that we can interpolate while(iter < maxit){ iter <- iter + 1 means <- matrix(0,II,J) ##contains temp of what we substract for (j in 1:(J-1)){ for (k in (j+1):J){ y <- mat[,j] - mat[,k] x <- (mat[,j] + mat[,k]) / 2 index <- c(order(x)[1], subset, order(-x)[1]) ##put endpoints in so we can interpolate xx <- x[index] yy <- y[index] aux <-loess(yy~xx, span=span, degree=1, weights=w, family=family.loess) aux <- predict(aux, data.frame(xx=x)) / J means[, j] <- means[, j] + aux means[, k] <- means[, k] - aux if (verbose) cat("Done with",j,"vs",k,"in iteration",iter,"\n") } } mat <- mat - means change <- max(colMeans((means[subset,])^2)) if(verbose) cat(iter, change,"\n") } if ((change > epsilon) & (maxit > 1)) warning(paste("No convergence after", maxit, "iterations.\n")) if(log.it) { return(2^mat) } else return(mat) } affy/R/normalize.qspline.R0000644000175200017520000001065714516003533016530 0ustar00biocbuildbiocbuildnormalize.AffyBatch.qspline <- function(abatch, type=c("together","pmonly","mmonly","separate"),...) { type <- match.arg(type) if (type == "together"){ Index <- unlist(indexProbes(abatch,"both")) intensity(abatch)[Index,] <- normalize.qspline(intensity(abatch)[Index,], ...) } else if (type == "pmonly"){ Index <- unlist(indexProbes(abatch,"pm")) intensity(abatch)[Index,] <- normalize.qspline(intensity(abatch)[Index,], ...) } else if (type == "mmonly"){ Index <- unlist(indexProbes(abatch,"mm")) intensity(abatch)[Index,] <- normalize.qspline(intensity(abatch)[Index,], ...) } else if (type == "separate"){ Index <- unlist(indexProbes(abatch,"pm")) intensity(abatch)[Index,] <- normalize.qspline(intensity(abatch)[Index,], ...) Index <- unlist(indexProbes(abatch,"mm")) intensity(abatch)[Index,] <- normalize.qspline(intensity(abatch)[Index,], ...) } #set.na.spotsd(listcel) normhisto <- vector("list", length=ncol(intensity(abatch))) ##need to use MIAME for this for (i in 1:length(abatch)) { normhisto[[i]] <- list(name="normalized by qspline") } attr(abatch, "normalization") <- normhisto return(abatch) } normalize.qspline <- function(x, target = NULL, samples = NULL, fit.iters = 5, min.offset = 5, spline.method = "natural", # c("fmm", "natural", "periodic") smooth = TRUE, spar = 0, # smoothing parameter p.min = 0, p.max = 1.0, incl.ends = TRUE, converge = FALSE, verbose = TRUE, na.rm = FALSE ){ if (is.null(target)) target <- exp(apply(log(x), 1, mean)) x.n <- dim(x)[1] m <- dim(x)[2] if (is.null(samples)) samples <- max(round(x.n/1000), 100) else if (samples < 1) samples <- round(samples * x.n) p <- (1:samples) / samples p <- p[ which(p <= p.max) & which(p >= p.min) ] samples <- length(p) k <- fit.iters if (na.rm==TRUE) y.n <- sum(!is.na(target)) else y.n <- length(target) py.inds <- as.integer(p * y.n) y.offset <- round(py.inds[1]/fit.iters) if (y.offset <= min.offset) { y.offset <- min.offset; k <- round(py.inds[1]/min.offset) } if (k <= 1) { warning("'k' found is non-sense. using default 'fit.iter'") k <- fit.iters } y.offset <- c(0, array(y.offset, (k-1))) y.order <- order(target) fx <- matrix(0, x.n,m) if(verbose==TRUE) print(paste("samples=",samples, "k=", k, "first=", py.inds[1])) for (i in 1:m) { # to handel NA values for each array if (na.rm==TRUE) x.valid <- which(!is.na(x[,i])) else x.valid <- 1:x.n x.n <- length(x.valid) px.inds <- as.integer(p * x.n) x.offset <- round(px.inds[1]/fit.iters) if (x.offset<=min.offset) { x.offset <- min.offset; k <- min(round(px.inds[1]/min.offset), k) } x.offset <- c(0, array(x.offset, (k-1))) x.order <- order(x[,i]) # NA's at the end (?) y.inds <- py.inds ## must be reset each iteration x.inds <- px.inds for (j in 1:k) { y.inds <- y.inds - y.offset[j] x.inds <- x.inds - x.offset[j] ty.inds <- y.inds tx.inds <- x.inds if (verbose==TRUE) print(paste("sampling(array=", i, "iter=", j, "off=", x.inds[1], -x.offset[j], y.inds[1], -y.offset[j], ")")) if (converge==TRUE) { ty.inds <- as.integer(c(1, y.inds)) tx.inds <- as.integer(c(1, x.inds)) if (j > 1) { ty.inds <- c(ty.inds, y.n) tx.inds <- c(tx.inds, x.n) } } qy <- target[y.order[ty.inds]] qx <- x[x.order[tx.inds],i] if (smooth==TRUE) { sspl <- smooth.spline(qx, qy, spar=spar) qx <- sspl$x qy <- sspl$y } fcn <- splinefun(qx, qy, method=spline.method) fx[x.valid,i] <- fx[x.valid,i] + fcn(x[x.valid,i])/k } if (na.rm==TRUE) { invalid <- which(is.na(x[,i])) fx[invalid,i] <- NA } } return(fx) } affy/R/normalize.quantiles.R0000644000175200017520000000755714516003533017067 0ustar00biocbuildbiocbuild################################################################## ## ## file: normalize.quantiles.R ## ## For a description of quantile normalization method see ## ## Bolstad, B. M., Irizarry R. A., Astrand, M, and Speed, T. P. (2003)(2003) ## A Comparison of Normalization Methods for High ## Density Oligonucleotide Array Data Based on Bias and Variance. ## Bioinformatics 19,2,pp 185-193 ## ## History ## Pre Aug 23, 2003 Two years worth of stuff ## Aug 23, 2003 - Added use.log2 to "robust", ## added ability to pass additional parameters ## to normalize.AffyBatch.Quantiles.robust ## changed pmonly parameters on functions ## so that it is now a string argument "type" ## the options are pmonly, mmonly, together, separate ## Jan 31, 2004 - put a check for an integer matrix and force coercision to ## doubles if required in normalize.quantiles ## Mar 13, 2005 - Modifications to normalize.quantiles.robust including removing ## approx.method which never got implemented. Making it a use a .Call() ## rather than a .C() ## ## Sep 20, 2006 - fix .Call in normalize.quantiles.robust ## May 20, 2007 - remove the functions that have been moved to preprocessCore ## ################################################################## normalize.AffyBatch.quantiles <- function(abatch,type=c("separate","pmonly","mmonly","together")) { type <- match.arg(type) if ((type == "pmonly")|(type == "separate")){ pms <- unlist(pmindex(abatch)) ## Change to faster computation of noNA - SDR 11/06/2003 ##noNA <- apply(intensity(abatch)[pms,,drop=FALSE],1,function(x) all(!is.na(x))) noNA <- rowSums(is.na(intensity(abatch)[pms,,drop=FALSE])) == 0 pms <- pms[noNA] intensity(abatch)[pms,] <- normalize.quantiles(intensity(abatch)[pms,,drop=FALSE ],copy=FALSE) } if((type == "mmonly") | (type == "separate")){ mms <- unlist(mmindex(abatch)) ## Change to faster computation of noNA - SDR 11/06/2003 ##noNA <- apply(intensity(abatch)[mms,,drop=FALSE],1,function(x) all(!is.na(x))) noNA <- rowSums(is.na(intensity(abatch)[mms,,drop=FALSE])) == 0 mms <- mms[noNA] intensity(abatch)[mms,] <- normalize.quantiles(intensity(abatch)[mms,,drop=FALSE ],copy=FALSE) } if (type == "together"){ pms <- unlist(indexProbes(abatch,"both")) intensity(abatch)[pms,] <- normalize.quantiles(intensity(abatch)[pms,,drop=FALSE ],copy=FALSE) } ##this is MIAME we need to decide how to do this properly. ##for (i in 1:length(abatch)) { ## history(abatch)[[i]]$name <- "normalized by quantiles" ##} return(abatch) } normalize.AffyBatch.quantiles.robust <- function(abatch, type=c("separate","pmonly","mmonly","together"),weights=NULL,remove.extreme=c("variance","mean","both","none"),n.remove=1,use.median=FALSE,use.log2=FALSE) { type <- match.arg(type) if ((type == "pmonly")|(type == "separate")){ pms <- unlist(pmindex(abatch)) intensity(abatch)[pms, ] <- normalize.quantiles.robust(intensity(abatch)[pms, ], copy=FALSE,weights=weights,remove.extreme,n.remove=n.remove,use.median=use.median,use.log2=use.log2) } if ((type == "mmonly")|(type == "separate")){ mms <- unlist(mmindex(abatch)) intensity(abatch)[mms, ] <- normalize.quantiles.robust(intensity(abatch)[mms, ],copy=FALSE,weights=weights,remove.extreme,n.remove=n.remove,use.median=use.median,use.log2=use.log2) } if (type == "together"){ pms <- unlist(indexProbes(abatch,"both")) intensity(abatch) <- normalize.quantiles.robust(intensity(abatch)[pms,,drop=FALSE ],copy=FALSE, weights=weights,remove.extreme=remove.extreme,n.remove=n.remove,use.median=use.median,use.log2=use.log2) } ##this is MIAME we need to decide how to do this properly. ##for (i in 1:length(abatch)) { ## history(abatch)[[i]]$name <- "normalized by quantiles" ##} return(abatch) } affy/R/pairs.AffyBatch.R0000644000175200017520000000257714516003533016024 0ustar00biocbuildbiocbuildpairs.AffyBatch <- function(x, panel=points, ..., transfo=I, main=NULL, oma=NULL, font.main = par("font.main"), cex.main = par("cex.main"), cex.labels = NULL, lower.panel=panel, upper.panel=NULL, diag.panel=NULL, #text.panel = textPanel, #label.pos = 0.5 + has.diag/3, font.labels = 1, row1attop = TRUE, gap = 1) { #label1 <- chipNames(x) #label2 <- unlist(lapply(history(x), function(z) z$name)) #textPanel <- function(x = 0.5, y = 0.5, txt, cex, font) { # text(x, y, txt, cex = cex, font = font) #} ##labels <- paste(sampleNames(x), unlist(lapply(history(x), function(z) if (is.null(z$name)) "" else z$name)), sep="\n") labels <- sampleNames(x) ##y <- matrix(intensity(x)[, , seq(along=x)], ncol=length(x)) y <- intensity(x) pairs(transfo(y), labels=labels, panel=panel, ..., main=main, oma=oma, font.main = font.main, cex.main = cex.main, lower.panel=lower.panel, upper.panel=upper.panel, diag.panel=diag.panel, #text.panel = text.panel, #label.pos = label.pos, cex.labels = cex.labels, font.labels = font.labels, row1attop = row1attop, gap = gap ) } affy/R/plot.ProbeSet.R0000644000175200017520000000060314516003533015544 0ustar00biocbuildbiocbuildplot.ProbeSet <- function(x, which=c("pm", "mm"), xlab="probes", type="l", ylim=NULL, ...) { which <- match.arg(which) if (which == "pm") f <- getMethod("pm", "ProbeSet") else f <- getMethod("mm", "ProbeSet") if (is.null(ylim)) ylim = range(c(f(x)), na.rm=TRUE) if (is.na(xlab)) xlab="probes" matplot(f(x), xlab=xlab, type=type, ylim=ylim, ...) } affy/R/plot.density.R0000644000175200017520000000272414516003533015506 0ustar00biocbuildbiocbuild# matdensity <- function(x, # ylab="density", xlab="x", type="l", plot=TRUE, # ...) { # x.density <- apply(mat, 2, density) # all.x <- do.call("cbind", lapply(x.density, function(x) x$x)) # all.y <- do.call("cbind", lapply(x.density, function(x) x$y)) # if (plot) # matplot(all.x, all.y, ylab=ylab, xlab=xlab, ...) # invisible(list(all.x=all.x, all.y=all.y)) # } plotDensity <- function(mat, ylab="density", xlab="x", type="l", col=1:6, na.rm = TRUE, ...) { x.density <- apply(mat, 2, density, na.rm = na.rm) all.x <- do.call(cbind, lapply(x.density, function(x) x$x)) all.y <- do.call(cbind, lapply(x.density, function(x) x$y)) matplot(all.x, all.y, ylab=ylab, xlab=xlab, type=type, col=col, ...) invisible(list(all.x=all.x, all.y=all.y)) } plotDensity.AffyBatch <- function(x, col=1:6, log=TRUE, which=c("pm","mm","both"), ylab="density", xlab=NULL, ...){ Index <- unlist(indexProbes(x, which=which)) x <- intensity(x)[Index, ,drop=FALSE] if(log){ x <- log2(x) if(is.null(xlab)) xlab <- "log intensity" } else if(is.null(xlab)) xlab <- "intensity" invisible(plotDensity(x, ylab=ylab, xlab=xlab, col=col, ...)) } affy/R/plotLocation.R0000644000175200017520000000113214516003533015511 0ustar00biocbuildbiocbuildplotLocation <- function(x, col="green", pch=22, ...) { if (is.list(x)) { x <- cbind(unlist(lapply(x, function(x) x[,1])), unlist(lapply(x, function(x) x[,2]))) } ## need to use nrow - x[,2] for correct y position. ## use image width to get nrow, which isn't ideal ## but follows assumption for this function that an ## image already exists nrow <- ceiling(par("usr")[4]) if(nrow == 1) stop(paste("\nYou must first generate an image of an array", "for this function to work!\n\n"), call. = FALSE) points(x[,1], nrow - x[,2] , pch=pch, col=col, ...) } affy/R/pmcorrect.mas.R0000644000175200017520000000221514516003533015622 0ustar00biocbuildbiocbuild############################ ##MPM Changed delta, Affy SADD states delta as 2e-20 pmcorrect.mas <- function (object, contrast.tau = 0.03, scale.tau = 10, delta = 2^(-20)) #function (object, contrast.tau = 0.03, scale.tau = 10, delta = 9.536743e-07) ########################### { all.pps.pm <- pm(object) all.pps.mm <- mm(object) diff <- log2(all.pps.pm) - log2(all.pps.mm) delta <- rep(delta, nrow(diff)) for (i in 1:ncol(diff)) { sb <- tukey.biweight(diff[, i]) pps.pm <- all.pps.pm[, i] pps.mm <- all.pps.mm[, i] pps.im <- pps.mm j <- (pps.mm >= pps.pm) & (sb > contrast.tau) pps.im[j] <- pps.pm[j]/2^sb j <- (pps.mm >= pps.pm) & (sb <= contrast.tau) pps.im[j] <- pps.pm[j]/2^(contrast.tau/(1 + (contrast.tau - sb)/scale.tau)) ######################### #MPM SADD Need to substract the PM-IM, I think this is the culprit pm.corrected <- apply(cbind(pps.pm-pps.im, delta), 1, max) #pm.corrected <- apply(cbind(pps.pm, pps.im, delta), 1, # max) ########################## diff[, i] <- pm.corrected } return(diff) } affy/R/ppset.ttest.R0000644000175200017520000000043114516003533015340 0ustar00biocbuildbiocbuildppset.ttest <- function(ppset, covariate, pmcorrect.fun = pmcorrect.pmonly, ...) { probes <- do.call(pmcorrect.fun, list(ppset)) my.ttest <- function(x) { y <- split(x, get(covariate)) t.test(y[[1]], y[[2]])$p.value } r <- apply(probes, 1, my.ttest) return(r) } affy/R/ppsetApply.R0000644000175200017520000000172614516003533015214 0ustar00biocbuildbiocbuildppsetApply <- function(abatch, FUN, genenames=NULL, ...) { if (! is(abatch, "AffyBatch")) stop("abatch must be inheriting from class AffyBatch") if (! is(FUN, "function")) stop("FUN must be a function") cdfenv <- getCdfInfo(abatch) if (is.null(genenames)) genenames <- ls(cdfenv) ## e1 <- new.env(parent = environment(FUN)) multiassign(names(pData(abatch)), pData(abatch), e1) environment(FUN) <- e1 ppset <- new("ProbeSet", pm=matrix(), mm=matrix()) r <- vector("list", length=length(genenames)) names(r) <- genenames for (i in seq(along=genenames)) { ## use mget to get NA when genenames[i] not found probes.i <- mget(genenames[i], envir = cdfenv, ifnotfound = NA)[[1]] if (all(is.na(probes.i))) next ppset@pm <- intensity(abatch)[probes.i[, 1], , drop=FALSE] ppset@mm <- intensity(abatch)[probes.i[, 2], , drop=FALSE] ppset@id <- genenames[i] r[[i]] <- FUN(ppset, ...) } return(r) } affy/R/read.affybatch.R0000644000175200017520000003340314516003533015711 0ustar00biocbuildbiocbuild############################################################# ## ## read.affybatch.R ## ## Adapted by B. M. Bolstad from read.affybatch in the affy ## package version 1.2. The goal is a faster, less memory hungry ## ReadAffy. To do this we will shunt more work off to ## the c code. ## ## History ## Jun 13-15 Intial version ## Jun 16 Verbose flag passed to C routine ## Jun 17 New method for checking header of first cel ## file. ## Jul 7 Added the function read.probematrix which ## reads in PM, MM or both into matrices ## Sep 28 changed name from read.affybatch2 to read.affybatch ## and cleaned up some old commented stuff ## Apr 13, 2004 - fixed problem in read.probematrix ## Nov 15, 2005 - add functionality to read the ## stddev values into the se.exprs slot (non-default behaviour) ## ## Jan 24, 2006 - JWM: added cdfname to allow for the use of non-standard mappings ## Mar 6, 2006 - change .Call to reference affyio. that is new location for parsing code ## Dec 12, 2006 - added checkCelFiles() to ensure all filenames are celfiles so unintended ## arguments don't get passed in via ... ## Apr 19, 2013 - JWM: added warning and error messages for Gene ST and Exon ST arrays ## Sept 26, 2013 - naked .Call() to affyio replaced ## ############################################################# read.affybatch <- function(..., filenames=character(0), phenoData=new("AnnotatedDataFrame"), description=NULL, notes="", compress = getOption("BioC")$affy$compress.cel, rm.mask = FALSE, rm.outliers=FALSE, rm.extra=FALSE, verbose = FALSE,sd=FALSE, cdfname = NULL) { auxnames <- unlist(list(...)) filenames <- c(filenames, auxnames) checkValidFilenames(filenames) n <- length(filenames) pdata <- pData(phenoData) ## try to read sample names form phenoData. if not there use CEL ## filenames if(dim(pdata)[1] != n) { ## if empty pdata filename are samplenames warning("Incompatible phenoData object. Created a new one.\n") samplenames <- sub("^/?([^/]*/)*", "", filenames) pdata <- data.frame(sample=1:n, row.names=samplenames) phenoData <- new("AnnotatedDataFrame", data=pdata, varMetadata=data.frame( labelDescription="arbitrary numbering", row.names="sample")) } else samplenames <- rownames(pdata) if (is.null(description)) { description <- new("MIAME") preproc(description)$filenames <- filenames preproc(description)$affyversion <- library(help=affy)$info[[2]][[2]][2] } if (length(notes)==0) notes(description) <- notes ## read the first file to see what we have if (verbose) cat(1, "reading",filenames[[1]],"...") headdetails <- read.celfile.header(as.character(filenames[[1]])) ##now we use the length dim.intensity <- headdetails[[2]] ##dim(intensity(cel)) ##and the cdfname as ref ref.cdfName <- headdetails[[1]] #cel@cdfName if(length(grep("gene1[01]st", cleancdfname(ref.cdfName))) == 1) warning(paste0("\n\nThe affy package can process data from the Gene ST 1.x series of arrays,\n", "but you should consider using either the oligo or xps packages, which are specifically\n", "designed for these arrays.\n\n"), call. = FALSE) if(length(grep("gene2[01]st|ex[1-2][0-1]st|hta20|mta10", cleancdfname(ref.cdfName))) == 1) stop(paste0("\n\nThe affy package is not designed for this array type.\n", "Please use either the oligo or xps package.\n\n"), call. = FALSE) scandates <- sapply(seq_len(length(filenames)), function(i) { sdate <- read.celfile.header(filenames[i], info = "full")[["ScanDate"]] if (is.null(sdate) ||length(sdate) == 0 ) NA_character_ else sdate }) protocol <- new("AnnotatedDataFrame", data=data.frame("ScanDate"=scandates, row.names=sampleNames(phenoData), stringsAsFactors=FALSE), dimLabels=c("sampleNames", "sampleColumns")) ## allow for non-standard cdfs if(is.null(cdfname)) cdfname <- ref.cdfName if (verbose) cat(paste("instantiating an AffyBatch (intensity a ", prod(dim.intensity), "x", length(filenames), " matrix)...", sep="")) if (verbose) cat("done.\n") ## Change sampleNames to be consistent with row.names of phenoData ## object exprs <- affyio::read_abatch(filenames, rm.mask, rm.outliers, rm.extra, ref.cdfName, dim.intensity[c(1,2)],verbose) colnames(exprs) <- samplenames #### this is where the code changes from the original read.affybatch. #### what we will do here is read in from the 1st to the nth CEL file if (!sd){ return(new("AffyBatch", exprs = exprs, ##se.exprs = array(NaN, dim=dim.sd), cdfName = cdfname, ##cel@cdfName, phenoData = phenoData, nrow = dim.intensity[2],##["Rows"], ncol = dim.intensity[1],##["Cols"], annotation = cleancdfname(cdfname, addcdf=FALSE), protocolData = protocol, description= description, notes = notes)) } else { return(new("AffyBatch", exprs = exprs, se.exprs = affyio::read_abatch_stddev(filenames, rm.mask, rm.outliers, rm.extra, ref.cdfName, dim.intensity[c(1,2)],verbose), cdfName = cdfname, ##cel@cdfName, phenoData = phenoData, nrow = dim.intensity[2],##["Rows"], ncol = dim.intensity[1],##["Cols"], annotation = cleancdfname(cdfname, addcdf=FALSE), protocolData = protocol, description= description, notes = notes)) } } ###################################################################################### read.probematrix <- function(..., filenames = character(0), phenoData = new("AnnotatedDataFrame"), description = NULL, notes = "", compress = getOption("BioC")$affy$compress.cel, rm.mask = FALSE, rm.outliers = FALSE, rm.extra = FALSE, verbose = FALSE,which="pm", cdfname = NULL){ auxnames <- unlist(list(...)) filenames <- c(filenames, auxnames) which <- match.arg(which,c("pm","mm","both")) if (verbose) cat(1, "reading", filenames[[1]], "to get header information\n") headdetails <- read.celfile.header(as.character(filenames[[1]])) ref.cdfName <- headdetails[[1]] cleaned.cdfName <- cleancdfname(ref.cdfName, addcdf = FALSE) ## Allow for usage of alternative cdfs if(is.null(cdfname)) Data <- new("AffyBatch", cdfName = ref.cdfName, annotation = cleaned.cdfName) else Data <- new("AffyBatch", cdfName = cdfname, annotation = cleaned.cdfName) cdfInfo <- as.list(getCdfInfo(Data)) cdfInfo <- cdfInfo[order(names(cdfInfo))] read.celfile.probeintensity.matrices(filenames = filenames, cdfInfo = cdfInfo, rm.mask = rm.mask, rm.outliers = rm.outliers, rm.extra = rm.extra, verbose = verbose, which = which) } list.celfiles <- function(...){ files <- list.files(...) return(files[grep("\\.[cC][eE][lL]\\.gz$|\\.[cC][eE][lL]$", files)]) } AllButCelsForReadAffy <- function(..., filenames=character(0), widget=getOption("BioC")$affy$use.widgets, celfile.path=NULL, sampleNames=NULL, phenoData=NULL, description=NULL){ ##first figure out filenames auxnames <- unlist(as.list(substitute(list(...)))[-1]) if (widget){ requireNamespace("tkWidgets") widgetfiles <- tkWidgets::fileBrowser( textToShow="Choose CEL files", testFun=tkWidgets::hasSuffix("[cC][eE][lL]|[cC][eE][lL].gz") ) } else{ widgetfiles <- character(0) } if(!is.null(celfile.path)){ auxnames <- file.path(celfile.path, auxnames) filenames <- file.path(celfile.path, filenames) } filenames <- c(filenames, auxnames, widgetfiles) if(length(filenames)==0){ if(is.null(celfile.path)) celfile.path <- getwd() filenames <- list.celfiles(celfile.path,full.names=TRUE) } if(length(filenames)==0) stop("No cel filennames specified and no cel files in specified directory:",celfile.path,"\n") if(is.null(sampleNames)){ sampleNames <- sub("^/?([^/]*/)*", "", filenames) } else{ if(length(sampleNames)!=length(filenames)){ warning("sampleNames not same length as filenames. Using filenames as sampleNames instead\n") sampleNames <- sub("^/?([^/]*/)*", "", filenames) } } chkSn <- function(filenames, samplenames){ fntest <- sub("^/?([^/]*/)*", "", filenames) if(all(fntest %in% samplenames)){ filenames <<- filenames[match(samplenames, fntest)] } else { warning(paste0("Mismatched phenoData and celfile names!\n\n", "Please note that the row.names of your phenoData ", "object should be identical to what you get from ", "list.celfiles()!\nOtherwise you are responsible for ", "ensuring that the ordering of your phenoData object ", "conforms to the ordering of the celfiles as they are ", "read into the AffyBatch!\nIf not, errors may ", "result from using the phenoData for subsetting or ", "creating linear models, etc.\n\n"), call. = FALSE) } } if(is.character(phenoData)) { ## if character, read file if(length(phenoData)!=1) stop(sprintf("'phenoData' must be of length 1, but is %d.", length(phenoData))) phenoData <- read.AnnotatedDataFrame(filename=phenoData) sampleNames <- sampleNames(phenoData) chkSn(filenames, sampleNames) } else if(is.data.frame(phenoData)) { ## if data.frame, coerce phenoData <- as(phenoData, "AnnotatedDataFrame") sampleNames <- sampleNames(phenoData) chkSn(filenames, sampleNames) } else if(is.null(phenoData)) { phenoData <- new("AnnotatedDataFrame", data = data.frame(sample=seq_along(sampleNames), row.names=sampleNames), varMetadata = data.frame(labelDescription="arbitrary numbering", row.names=names(pData))) } else if (!is(phenoData, "AnnotatedDataFrame")) { stop(sprintf("'phenoData' must be of class 'AnnotatedDataFrame', but is %s.", class(phenoData))) } ##get MIAME information if(is.character(description)){ description <- read.MIAME(filename=description,widget=FALSE) } else{ if (! is(description, "MIAME")) { description <- new("MIAME") } } ##MIAME stuff description@preprocessing$filenames <- filenames description@preprocessing$affyversion <- library(help=affy)$info[[2]][[2]][2] return(list(filenames = filenames, phenoData = phenoData, sampleNames = sampleNames, description = description)) } ###this is user friendly wrapper for read.affybatch ReadAffy <- function(..., filenames=character(0), widget=getOption("BioC")$affy$use.widgets, compress=getOption("BioC")$affy$compress.cel, celfile.path=NULL, sampleNames=NULL, phenoData=NULL, description=NULL, notes="", rm.mask=FALSE, rm.outliers=FALSE, rm.extra=FALSE, verbose=FALSE,sd=FALSE, cdfname = NULL) { l <- AllButCelsForReadAffy(..., filenames=filenames, widget=widget, celfile.path=celfile.path, sampleNames=sampleNames, phenoData=phenoData, description=description) ##and now we are ready to read cel files ret <- read.affybatch(filenames=l$filenames, phenoData=l$phenoData, description=l$description, notes=notes, compress=compress, rm.mask=rm.mask, rm.outliers=rm.outliers, rm.extra=rm.extra, verbose=verbose,sd=sd,cdfname=cdfname) sampleNames(ret) <- l$sampleNames return(ret) } checkValidFilenames <- function(filenames) { ## Returns TRUE if filenames is a character vector containing ## paths to files that exist (directories don't count). ## A suitable error message is printed via stop() if invalid ## file names are encountered. if (!is.character(filenames)) stop(strwrap(paste("file names must be specified using a character", "vector, not a", sQuote(typeof(filenames)))), call.=FALSE) if (length(filenames) == 0) stop("no file names provided") if (any(sapply(filenames, nchar) < 1)) stop("empty file names are not allowed") finfo <- file.info(filenames) whBad <- sapply(finfo[["isdir"]], function(x) !identical(FALSE, x)) if (any(whBad)) { msg <- paste("the following are not valid files:\n", paste(" ", filenames[whBad], collapse="\n")) stop(msg, call.=FALSE) } TRUE } affy/R/rma.R0000644000175200017520000000414614516003533013631 0ustar00biocbuildbiocbuild###################################################### # # rma - RMA interface to c code # # the RMA method implemented in c code # # this code serves as interface to the c code. # currently # implemented (version 0.25) background correction # # Background correction code has been added. # # note this function does not leave the supplied # AffyBatch unchanged if you select DESTRUCTIVE=TRUE. this is # for memory purposes but can be quite # dangerous if you are not careful. Use destructive=FALSE if this is # deemed likely to be a problem. # # UPDATE: note that the affybatch is now not affected if you use # destructive=TRUE and you might actually save a little memory. # the destructive refers only to Plobs, which would be destroyed. # # History # # Feb 22, 2004 - activated subset. In is now possible to # do the entire RMA procedure using a subset of probesets # # Oct 26, 2007 = makesure verbosity flag is correctly passed down to C-level routines # # Oct 28, 2007 MM are no longer passed to the C code # # Jul 2, 2008 - change how probeNames (which is really probe/row indexing) is passed down to # c code # # ######################################################## rma <- function(object,subset=NULL, verbose=TRUE, destructive = TRUE,normalize=TRUE,background=TRUE,bgversion=2,...){ rows <- length(probeNames(object,subset)) cols <- length(object) if (is.null(subset)){ ngenes <- length(geneNames(object)) } else { ngenes <- length(subset) } pNList <- probeNames(object,subset) pNList <- split(0:(length(pNList) -1), pNList) if (destructive){ exprs <- .Call("rma_c_complete",pm(object,subset), pNList, ngenes, normalize, background, bgversion, verbose, PACKAGE="affy") } else { exprs <- .Call("rma_c_complete_copy", pm(object,subset), pNList, ngenes, normalize, background, bgversion, verbose, PACKAGE="affy") } colnames(exprs) <- sampleNames(object) new("ExpressionSet", phenoData = phenoData(object), annotation = annotation(object), protocolData = protocolData(object), experimentData = experimentData(object), exprs = exprs) } affy/R/summary.R0000644000175200017520000000344114516003533014544 0ustar00biocbuildbiocbuild###these are summary functions they take matrices of probes x chips ###and return expression and se (when applicable) ##DEBUG: appending the se to the expression values in a same vector ## is too much hackish (I think)... we need to think about something ## better avdiff <- function(x,constant=3){ e <- apply(x,2,function(y){ o <- order(y) yy <- y[-c(o[1],o[length(y)])] #take out biggest and smallest if(length(yy)<2) # SK, some genes have only one probe mean(y) else mean(y[abs(y-mean(yy))detecting normalization methods from naming convention\n") ## this could move into the respective methods of AffyBatch later assign("normalize.AffyBatch.methods", sub("^normalize\\.AffyBatch\\.", "", grep("^normalize.AffyBatch", all.affy, value = TRUE)), envir=env) } .initExpression <- function(all.affy, env) { if (debug.affy123) cat("-->detecting expression value methods from naming convention\n") ## the first one is deprecated (well... "should be"...) vals <- sub("^generateExprVal\\.method\\.", "", grep("^generateExprVal.method", all.affy, value = TRUE)) assign("generateExprSet.methods", vals, envir=env) assign("express.summary.stat.methods", vals, envir=env) } .initBackgroundCorrect <- function(all.affy, env) { if (debug.affy123) cat("-->detecting background correction methods from naming convention\n") start <- nchar("bg.correct.") assign("bgcorrect.methods", sub("^bg\\.correct\\.", "", grep("^bg.correct", all.affy, value = TRUE)), envir=env) } .initPmCorrect <- function(all.affy, env) { if (debug.affy123) cat("-->detecting pm correction methods from naming convention\n") assign("pmcorrect.methods", sub("^pmcorrect\\.", "", grep("^pmcorrect", all.affy, value = TRUE)), envir=env) } .setAffyOptions <- function(affy.opt=NA) { if (! any(is.na(affy.opt))) { if (class(affy.opt) != "BioCPkg") stop("obviously invalid package options !") BioC <- getOption("BioC") BioC$affy <- affy.opt options("BioC"=BioC) return() } ## add affy specific options ## (not unlike what is done in 'Biobase') if (is.null(getOption("BioC"))) { BioC <- list() class(BioC) <- "BioCOptions" options("BioC"=BioC) } probesloc.first <- list(what="environment", where=.GlobalEnv) probesloc.second <- list(what="libPath", where=NULL) probesloc.third <- list(what="data", where="affy") probesloc.fourth <- list(what="bioC", where=.libPaths()[1]) ## default for the methods bgcorrect.method <- "mas" normalize.method <- "quantiles" pmcorrect.method <- "pmonly" summary.method <- "liwong" affy.opt <- list(compress.cdf=FALSE, compress.cel=FALSE, use.widgets=FALSE, probesloc = list(probesloc.first, probesloc.second, probesloc.third, probesloc.fourth), bgcorrect.method = bgcorrect.method, normalize.method = normalize.method, pmcorrect.method = pmcorrect.method, summary.method = summary.method, xy.offset = 0 ## this one is for temporary compatibility ) class(affy.opt) <- "BioCPkg" BioC <- getOption("BioC") BioC$affy <- affy.opt options("BioC"=BioC) ## --- } .affyInternalEnv <- NULL .onLoad <- function(libname, pkgname) { # where <- match(paste("package:", pkgname, sep=""), search()) all.affy <- ls(environment(sys.function())) ##a place to store some variables that need to be accessed .affyInternalEnv <<- new.env(parent=emptyenv()) .initNormalize(all.affy, .affyInternalEnv) .initExpression(all.affy, .affyInternalEnv) .initBackgroundCorrect(all.affy, .affyInternalEnv) .initPmCorrect(all.affy, .affyInternalEnv) .setAffyOptions() if(.Platform$OS.type == "windows" && interactive() && .Platform$GUI == "Rgui"){ addVigs2WinMenu("affy") } } affy/README.md0000644000175200017520000000063714516003533014006 0ustar00biocbuildbiocbuild[](https://bioconductor.org/) **affy** is an R/Bioconductor package conaining methods for Affymetrix oligonucleotide arrays. See https://bioconductor.org/packages/affy for more information including how to install the release version of the package (please refrain from installing directly from GitHub). affy/aclocal.m40000755000175200017520000000113414516003533014363 0ustar00biocbuildbiocbuild## ## Try finding zlib library and headers ## ## R_ZLIB() ## AC_DEFUN([R_ZLIB], [ have_zlib=no AC_CHECK_LIB(z, main, [ AC_CHECK_HEADER(zlib.h, [ AC_MSG_CHECKING([if zlib version >= 1.1.3]) AC_TRY_RUN([ #include "confdefs.h" #include #include int main() { #ifdef ZLIB_VERSION return(strcmp(ZLIB_VERSION, "1.1.3") < 0); #else return(1); #endif }], [AC_MSG_RESULT([yes]) have_zlib=yes], AC_MSG_RESULT([no]), AC_MSG_RESULT([no])) ]) ]) if test "${have_zlib}" = yes; then AC_DEFINE(HAVE_ZLIB) LIBS='-lz '$LIBS fi ]) affy/build/0000755000175200017520000000000014516022632013621 5ustar00biocbuildbiocbuildaffy/build/vignette.rds0000644000175200017520000000050414516022632016157 0ustar00biocbuildbiocbuildR]O0kl,nMb|hڒ҉{م =bH˽ܞ{i}0,ñMòf For some (mysterious) reasons, save(..., ascii=TRUE) is behaving wrong on my linux box at home. I saved it with ascii=FALSE (portability could be .....) LG ###why not have a slot in cel.container denoting the default Cdf ###then change all functions so that one does not need to pass Cdf ###with this and some methods we maybe able to get rid of Plobs entirely ###maybe this is a version 2.0 task. ----> Yep... mentioned in the file TODO for some time now... Yo, Laurentz takin' the mike now: - fixed a problem with CDF.example.RData - updated the documentation for the normalize functions (should keep 'R CMD check happy') - introduced a generic method 'normalize.methods' (Plob and Cel.container) - variables related to normalize.methods (normalize.Cel.container.methods and normalize.Plob.methods) are created. - changed the methods names 'normalize.Cel.XYZ' to 'normalize.Cel.container.XYZ' (more consistent). Documentation shoudlhave been updated accordingly. - removed image.Cel.Rd - fixed the method image for Cel-class. Added better ways of asking the function to mask areas. updated the documentation accordingly. - did a bit more with affy.Rnw (in inst/doc/). Now it goes through Sweave ok. - update the file 'TODO' still talkin'...: - cleaned read_cdffile.c (no more warnings) - made something usable for the documentation of 'normalize' (hopefully) - various fixes in the documentation - Did a bit more with affy.Rnw - the example in generateExprSet-methods.Rd is commented out... seems to work on the command line but fails through 'R CMD check' (??!?!!!?) - the 'normalize' part being done (apart of possible bugs), we reached the version number 0.8.0 - started to finish the part about generateExprSet - accessor function sd in Cel moved to spotsd affy/cleanup0000755000175200017520000000005014516022632014072 0ustar00biocbuildbiocbuild#! /bin/sh rm -f config.* src/Makevars affy/configure0000755000175200017520000007614214516022632014443 0ustar00biocbuildbiocbuild#! /bin/sh # Guess values for system-dependent variables and create Makefiles. # Generated automatically using autoconf version 2.13 # Copyright (C) 1992, 93, 94, 95, 96 Free Software Foundation, Inc. # # This configure script is free software; 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"s%^%$ac_given_srcdir/%" -e "s%:% $ac_given_srcdir/%g"` sed -e "$ac_comsub s%@configure_input@%$configure_input%g s%@srcdir@%$srcdir%g s%@top_srcdir@%$top_srcdir%g " $ac_file_inputs | (eval "$ac_sed_cmds") > $ac_file fi; done rm -f conftest.s* EOF cat >> $CONFIG_STATUS <> $CONFIG_STATUS <<\EOF exit 0 EOF chmod +x $CONFIG_STATUS rm -fr confdefs* $ac_clean_files test "$no_create" = yes || ${CONFIG_SHELL-/bin/sh} $CONFIG_STATUS || exit 1 affy/configure.in0000755000175200017520000000107114516003533015034 0ustar00biocbuildbiocbuilddnl dnl Configuration things for affyR. dnl (http://www.cbs.dtu.dk/laurent/download/affyR/ dnl What is below (and in the other configuration fiels dnl was taken from different configuration scripts for R version 1.3.0. dnl dnl Acknowledgments: The author(s) of the R configure scripts, Kurt Hornik for the tip with autoconf. dnl dnl Laurent 2001 AC_INIT("DESCRIPTION") dnl dnl Are things (still) the same ? dnl (taken from the 'writing R extensions manual') dnl dnl 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/0 m^؛hoFwmc:f#:_s}~ϯԑ|~o)x7@}~o->YS*ve+KG~b~'G?ߕ2r=uGQuOGqpԽQ(n TWI4q.w=.cyӚɥq.x+<?m6KJ %9 9LE#usGstgsNru\gu9s9i{NyZi^&} ]gu&0affy/data/mapCdfName.rda0000644000175200017520000000032714516003533016117 0ustar00biocbuildbiocbuild r0b```b`RL@& `d`\)i~ @( RWZ[ 0640O40]s2SAaGaq!T5hTl`pm(Dilution)) ################################################### ### code chunk number 15: affy.Rnw:515-517 ################################################### gn <- geneNames(Dilution) pm(Dilution, gn[100]) ################################################### ### code chunk number 16: affy.Rnw:531-532 ################################################### hist(Dilution[,1:2]) ##PM histogram of arrays 1 and 2 ################################################### ### code chunk number 17: affy.Rnw:548-550 (eval = FALSE) ################################################### ## par(mfrow=c(2,2)) ## image(Dilution) ################################################### ### code chunk number 18: affy.Rnw:566-568 ################################################### par(mfrow=c(1,1)) boxplot(Dilution, col=c(2,3,4)) ################################################### ### code chunk number 19: affy.Rnw:591-593 ################################################### deg <- AffyRNAdeg(Dilution) names(deg) ################################################### ### code chunk number 20: affy.Rnw:597-598 ################################################### summaryAffyRNAdeg(deg) ################################################### ### code chunk number 21: affy.Rnw:605-606 ################################################### plotAffyRNAdeg(deg) ################################################### ### code chunk number 22: affy.Rnw:620-621 ################################################### Dilution.normalized <- normalize(Dilution) ################################################### ### code chunk number 23: affy.Rnw:667-671 ################################################### gn <- featureNames(Dilution) ps <- probeset(Dilution, gn[1:2]) #this is what i should be using: ps show(ps[[1]]) ################################################### ### code chunk number 24: affy.Rnw:687-689 ################################################### mylocation <- list("1000_at"=cbind(pm=c(1,2,3),mm=c(4,5,6)), "1001_at"=cbind(pm=c(4,5,6),mm=c(1,2,3))) ################################################### ### code chunk number 25: affy.Rnw:696-698 ################################################### ps <- probeset(Dilution, genenames=c("1000_at","1001_at"), locations=mylocation) ################################################### ### code chunk number 26: affy.Rnw:702-706 ################################################### pm(ps[[1]]) mm(ps[[1]]) pm(ps[[2]]) mm(ps[[2]]) ################################################### ### code chunk number 27: affy.Rnw:725-737 ################################################### data(SpikeIn) ##SpikeIn is a ProbeSets pms <- pm(SpikeIn) mms <- mm(SpikeIn) ##pms follow concentration par(mfrow=c(1,2)) concentrations <- matrix(as.numeric(sampleNames(SpikeIn)),20,12,byrow=TRUE) matplot(concentrations,pms,log="xy",main="PM",ylim=c(30,20000)) lines(concentrations[1,],apply(pms,2,mean),lwd=3) ##so do mms matplot(concentrations,mms,log="xy",main="MM",ylim=c(30,20000)) lines(concentrations[1,],apply(mms,2,mean),lwd=3) ################################################### ### code chunk number 28: affy.Rnw:771-773 ################################################### cat("HG_U95Av2 is",cleancdfname("HG_U95Av2"),"\n") cat("HG-133A is",cleancdfname("HG-133A"),"\n") ################################################### ### code chunk number 29: affy.Rnw:777-778 ################################################### cat("HG_U95Av2 is",cleancdfname("HG_U95Av2",addcdf=FALSE),"\n") ################################################### ### code chunk number 30: affy.Rnw:785-788 ################################################### data(cdfenv.example) ls(cdfenv.example)[1:5] get(ls(cdfenv.example)[1],cdfenv.example) ################################################### ### code chunk number 31: affy.Rnw:799-802 ################################################### print(Dilution@cdfName) myenv <- getCdfInfo(Dilution) ls(myenv)[1:5] ################################################### ### code chunk number 32: affy.Rnw:810-813 ################################################### Index <- pmindex(Dilution) names(Index)[1:2] Index[1:2] ################################################### ### code chunk number 33: affy.Rnw:817-818 ################################################### pmindex(Dilution, genenames=c("1000_at","1001_at")) ################################################### ### code chunk number 34: affy.Rnw:822-823 ################################################### mmindex(Dilution, genenames=c("1000_at","1001_at")) ################################################### ### code chunk number 35: affy.Rnw:826-829 ################################################### indexProbes(Dilution, which="pm")[1] indexProbes(Dilution, which="mm")[1] indexProbes(Dilution, which="both")[1] ################################################### ### code chunk number 36: affy.Rnw:841-844 ################################################### opt <- getOption("BioC") affy.opt <- opt$affy print(names(affy.opt)) ################################################### ### code chunk number 37: affy.Rnw:848-853 ################################################### opt <- getOption("BioC") affy.opt <- opt$affy affy.opt$normalize.method <- "constant" opt$affy <- affy.opt options(BioC=opt) ################################################### ### code chunk number 38: affy.Rnw:859-864 ################################################### opt <- getOption("BioC") affy.opt <- opt$affy affy.opt$compress.cel <- TRUE opt$affy <- affy.opt options(BioC=opt) affy/inst/doc/affy.Rnw0000644000175200017520000010542014516003533015662 0ustar00biocbuildbiocbuild% -*- mode: noweb; noweb-default-code-mode: R-mode; -*- %\VignetteIndexEntry{1. Primer} %\VignetteKeywords{Preprocessing, Affymetrix} %\VignetteDepends{affy} %\VignettePackage{affy} %documentclass[12pt, a4paper]{article} \documentclass[12pt]{article} \usepackage{amsmath} \usepackage{hyperref} \usepackage[authoryear,round]{natbib} \textwidth=6.2in \textheight=8.5in %\parskip=.3cm \oddsidemargin=.1in \evensidemargin=.1in \headheight=-.3in \newcommand{\scscst}{\scriptscriptstyle} \newcommand{\scst}{\scriptstyle} \newcommand{\Rfunction}[1]{{\texttt{#1}}} \newcommand{\Robject}[1]{{\texttt{#1}}} \newcommand{\Rpackage}[1]{{\textit{#1}}} \author{Laurent Gautier, Rafael Irizarry, Leslie Cope, and Ben Bolstad} \begin{document} \title{Description of affy} \maketitle \tableofcontents \section{Introduction} The \Rpackage{affy} package is part of the BioConductor\footnote{\url{http://bioconductor.org/}} project. It is meant to be an extensible, interactive environment for data analysis and exploration of Affymetrix oligonucleotide array probe level data. The software utilities provided with the Affymetrix software suite summarizes the probe set intensities to form one {\it expression measure} for each gene. The expression measure is the data available for analysis. However, as pointed out by \cite{li:wong:2001a}, much can be learned from studying the individual probe intensities, or as we call them, the {\it probe level data}. This is why we developed this package. The package includes plotting functions for the probe level data useful for quality control, RNA degradation assessments, different probe level normalization and background correction procedures, and flexible functions that permit the user to convert probe level data to expression measures. The package includes utilities for computing expression measures similar to MAS 4.0's AvDiff \citep{affy4}, MAS 5.0's signal \citep{affy5}, DChip's MBEI \citep{li:wong:2001a}, and RMA \citep{iriz:etal:2003}. We assume that the reader is already familiar with oligonucleotide arrays and with the design of the Affymetrix GeneChip arrays. If you are not, we recommend the Appendix of the Affymetrix MAS manual \cite{affy4,affy5}. The following terms are used throughout this document: \begin{description} \item[probe] oligonucleotides of 25 base pair length used to probe RNA targets. \item[perfect match] probes intended to match perfectly the target sequence. \item[$PM$] intensity value read from the perfect matches. \item[mismatch] the probes having one base mismatch with the target sequence intended to account for non-specific binding. \item[$MM$] intensity value read from the mis-matches. \item[probe pair] a unit composed of a perfect match and its mismatch. \item[affyID] an identification for a probe set (which can be a gene or a fraction of a gene) represented on the array. \item[probe pair set] $PM$s and $MM$s related to a common {\it affyID}. \item[{\it CEL} files] contain measured intensities and locations for an array that has been hybridized. \item[{\it CDF} file] contain the information relating probe pair sets to locations on the array. \end{description} Section \ref{whatsnew} describes the main differences between version 1.5 and this version (1.6). Section \ref{sec:get.started} describes a quick way of getting started and getting expression measures. Section \ref{qc} describes some quality control tools. Section \ref{s1.4} describes normalization routines. Section \ref{classes} describes the different classes in the package. \ref{sec:probesloc} describes our strategy to map probe locations to probe set membership. Section \ref{configure.options} describes how to change the package's default options. Section \ref{whatwasnew} describes earlier changes. %%%make sure to change this when we get a publication about version 2. {\bf Note:} If you use this package please cite \cite{gaut:cope:bols:iriz:2003} and/or \cite{iriz:gaut:cope:2003}. \section{Changes for affy in BioC 1.8 release} \label{whatsnew} There were relatively few changes. \begin{itemize} \item MAplot now accepts the argument \Rfunction{plot.method} which can be used to call smoothScatter. \item \Rfunction{normalize.quantiles.robust} has had minor changes. \item \Rfunction{ReadAffy} can optionally return the SD values stored in the cel file. \item The C parsing code has been moved to the \Rpackage{affyio} package, which is now a dependency of the affy package. This change should be transparent to users as \Rpackage{affyio} will be automatically loaded when affy is loaded. \item Added a cdfname argument to \Rfunction{justRMA} and \Rfunction{ReadAffy} to allow for the use of alternative cdf packages. \end{itemize} \section{Getting Started: From probe level data to expression values} \label{sec:get.started} The first thing you need to do is {\bf load the package}. \begin{Sinput} R> library(affy) ##load the affy package \end{Sinput} <>= library(affy) @ This release of the \Rpackage{affy} package will automatically download the appropriate cdf environment when you require it. However, if you wish you may download and install the cdf environment you need from \url{http://bioconductor.org/help/bioc-views/release/data/annotation/} manually. If there is no cdf environment currently built for your particular chip and you have access to the CDF file then you may use the \Rpackage{makecdfenv} package to create one yourself. To make the cdf packaes, Microsoft Windows users will need to use the tools described in \url{http://www.murdoch-sutherland.com/Rtools/}. \subsection{Quick start} If all you want is to go from probe level data ({\it Cel} files) to expression measures here are some quick ways. If you want is RMA, the quickest way of reading in data and getting expression measures is the following: \begin{enumerate} \item Create a directory, move all the relevant {\it CEL} files to that directory \item If using linux/unix, start R in that directory. \item If using the Rgui for Microsoft Windows make sure your working directory contains the {\it Cel} files (use ``File -> Change Dir'' menu item). \item Load the library. \begin{Sinput} R> library(affy) ##load the affy package \end{Sinput} \item Read in the data and create an expression, using RMA for example. \begin{Sinput} R> Data <- ReadAffy() ##read data in working directory R> eset <- rma(Data) \end{Sinput} \end{enumerate} Depending on the size of your dataset and on the memory available to your system, you might experience errors like `Cannot allocate vector \ldots'. An obvious option is to increase the memory available to your R process (by adding memory and/or closing external applications\footnote{UNIX-like systems users might also want to check {\it ulimit} and/or compile {\bf R} and the package for 64 bits when possible.}. An another option is to use the function \Rfunction{justRMA}. \begin{Sinput} R> eset <- justRMA() \end{Sinput} This reads the data and performs the `RMA' way to preprocess them at the {\it C} level. One does not need to call \verb+ReadAffy+, probe level data is never stored in an AffyBatch. \verb+rma+ continues to be the recommended function for computing RMA. The \Rfunction{rma} function was written in C for speed and efficiency. It uses the expression measure described in \cite{iriz:etal:2003}. For other popular methods use \Rfunction{expresso} instead of \Rfunction{rma} (see Section \ref{expresso}). For example for our version of MAS 5.0 signal uses expresso (see code). To get mas 5.0 you can use \begin{Sinput} R> eset <- mas5(Data) \end{Sinput} which will also normalize the expression values. The normalization can be turned off through the \verb+normalize+ argument. In all the above examples, the variable \Robject{eset} is an object of class \Robject{ExpressionSet} described in the Biobase vignette. Many of the packages in BioConductor work on objects of this class. See the \Rpackage{genefilter} and \Rpackage{geneplotter} packages for some examples. If you want to use some other analysis package, you can write out the expression values to file using the following command: \begin{Sinput} R> write.exprs(eset, file="mydata.txt") \end{Sinput} \subsection{Reading CEL file information} The function \Rfunction{ReadAffy} is quite flexible. It lets you specify the filenames, phenotype, and MIAME information. You can enter them by reading files (see the help file) or widgets (you need to have the tkWidgets package installed and working). \begin{Sinput} R> Data <- ReadAffy(widget=TRUE) ##read data in working directory \end{Sinput} This function call will pop-up a file browser widget, see Figure \ref{fig:widget.filechooser}, that provides an easy way of choosing cel files. \newpage \begin{figure}[htbp] \begin{center} \includegraphics{widgetfilechooser} \caption{\label{fig:widget.filechooser}Graphical display for selecting {\it CEL} files. This widget is part of the {\it tkWidgets} package. (function written by Jianhua (John) Zhang). } \end{center} \end{figure} Next, a widget (not shown) permits the user to enter the \verb+phenoData+. %%See Figure \ref{fig:widget.pd}. %% \begin{figure}[htbp] %% \begin{center} %% \begin{tabular}{c} %% \includegraphics{numcovariates}\\ %% \includegraphics{namecovariates}\\ %% \includegraphics{assigncovariates} %% \end{tabular} %% \caption{\label{fig:widget.pd}Graphical display for entering phenoData %% This widget is part %% of the {\it tkWidgets} package.} %% % (functions written by Majnu John.} %% \end{center} %% \end{figure} Finally the a widget is presented for the user to enter MIAME information. %%Seen in Figure \ref{fig:widget.tkMIAME}. %% \begin{figure}[htbp] %% \begin{center} %% \includegraphics[width=0.5\textwidth]{widgettkMIAME} %% \caption{\label{fig:widget.tkMIAME}Graphical display for entering {\it %% MIAME} informations. This widget is part of the {\it tkWidgets} %% package.} %% % (function written by Majnu John).} %% \end{center} %% \end{figure} Notice that it is not necessary to use widgets to enter this information. Please read the help file for more information on how to read it from flat files or to enter it programmatically. The function \Rfunction{ReadAffy} is a wrapper for the functions \Rfunction{read.affybatch}, \Rfunction{tkSampleNames}, \Rfunction{read.AnnotatedDataFrame}, and \Rfunction{read.MIAME}. The function \Rfunction{read.affybatch} has some nice feature that make it quite flexible. For example, the \verb+compression+ argument permit the user to read compressed {\it CEL} files. The argument {\it compress} set to {\it TRUE} will inform the readers that your files are compressed and let you read them while they remain compressed. The compression formats {\it zip} and {\it gzip} are known to be recognized. A comprehensive description of all these options is found in the help file: \begin{Sinput} R> ?read.affybatch R> ?read.AnnotatedDataFrame R> ?read.MIAME \end{Sinput} \subsection{Expression measures} The most common operation is certainly to convert probe level data to expression values. Typically this is achieved through the following sequence: \begin{enumerate} \item reading in probe level data. \item background correction. \item normalization. \item probe specific background correction, e.g. subtracting $MM$. \item summarizing the probe set values into one expression measure and, in some cases, a standard error for this summary. \end{enumerate} We detail what we believe is a good way to proceed below. As mentioned the function \Rfunction{expresso} provides many options. For example, \begin{Sinput} R> eset <- expresso(Dilution, normalize.method="qspline", bgcorrect.method="rma",pmcorrect.method="pmonly", summary.method="liwong") \end{Sinput} This will store expression values, in the object \Robject{eset}, as an object of class \Robject{ExpressionSet} (see the \Rpackage{Biobase} package). You can either use R and the BioConductor packages to analyze your expression data or if you rather use another package you can write it out to a tab delimited file like this \begin{Sinput} R> write.exprs(eset, file="mydata.txt") \end{Sinput} In the \verb+mydata.txt+ file, row will represent genes and columns will represent samples/arrays. The first row will be a header describing the columns. The first column will have the {\it affyID}s. The \Rfunction{write.exprs} function is quite flexible on what it writes (see the help file). \subsubsection{expresso} \label{expresso} The function \Rfunction{expresso} performs the steps background correction, normalization, probe specific correction, and summary value computation. We now show this using an \Robject{AffyBatch} included in the package for examples. The command \verb+data(Dilution)+ is used to load these data. Important parameters for the expresso function are: \begin{description} \item[bgcorrect.method]. The background correction method to use. The available methods are <<>>= bgcorrect.methods() @ \item[normalize.method]. The normalization method to use. The available methods can be queried by using \verb+normalize.methods+. <<>>= library(affydata) data(Dilution) ##data included in the package for examples normalize.methods(Dilution) @ \item[pmcorrect.method] The method for probe specific correction. The available methods are <<>>= pmcorrect.methods() @ \item[summary.method]. The summary method to use. The available methods are <<>>= express.summary.stat.methods() @ Here we use \Rfunction{mas} to refer to the methods described in the Affymetrix manual version 5.0. \item[widget] Making the \verb+widget+ argument \verb+TRUE+, will let you select missing parameters (like the normalization method, the background correction method or the summary method). Figure \ref{fig:expressochooser} shows the widget for the selection of preprocessing methods for each of the steps. \begin{Sinput} R> expresso(Dilution, widget=TRUE) \end{Sinput} \begin{figure}[htbp] \begin{center} \includegraphics[width=0.5\textwidth]{EWSnap} \caption{\label{fig:expressochooser}Graphical display for selecting expresso methods.} \end{center} \end{figure} \end{description} There is a separate vignette {\bf affy: Built-in Processing Methods} which explains in more detail what each of the preprocessing options does. \subsubsection{MAS 5.0} To obtain expression values that correspond to those from MAS 5.0, use \Rfunction{mas5}, which wraps \Rfunction{expresso} and \Rfunction{affy.scalevalue.exprSet}. <<>>= eset <- mas5(Dilution) @ To obtain MAS 5.0 presence calls you can use the \verb+mas5calls+ method. <<>>= Calls <- mas5calls(Dilution) @ This returns an \verb+ExpressionSet+ object containing P/M/A calls and their associated Wilcoxon p-values. \subsubsection{Li and Wong's MBEI (dchip)} To obtain our version of Li and Wong's MBEI one can use \begin{Sinput} R> eset <- expresso(Dilution, normalize.method="invariantset", bg.correct=FALSE, pmcorrect.method="pmonly",summary.method="liwong") \end{Sinput} This gives the current $PM$-only default. The reduced model (previous default) can be obtained using \verb+pmcorrect.method="subtractmm"+. \subsubsection{C implementation of RMA} One of the quickest ways to compute expression using the \Rpackage{affy} package is to use the \Rfunction{rma} function. We have found that this method allows a user to compute the RMA expression measure in a matter of minutes for datasets that may have taken hours in previous versions of \Rpackage{affy}. The function serves as an interface to a hard coded C implementation of the RMA method \citep{iriz:etal:2003}. Generally, the following would be sufficient to compute RMA expression measures: <<>>= eset <- rma(Dilution) @ Currently the \Rfunction{rma} function implements RMA in the following manner \begin{enumerate} \item Probe specific correction of the PM probes using a model based on observed intensity being the sum of signal and noise \item Normalization of corrected PM probes using quantile normalization \citep{bols:etal:2003} \item Calculation of Expression measure using median polish. \end{enumerate} The \Rfunction{rma} function is likely to be improved and extended in the future as the RMA method is fine-tuned. \newpage \section{Quality Control through Data Exploration} \label{qc} For the users convenience we have included the \verb+Dilution+ sample data set: <<>>= Dilution @ This will create the \verb+Dilution+ object of class \Robject{AffyBatch}. \Rfunction{print} (or \Rfunction{show}) will display summary information. These objects represent data from one experiment. The \Robject{AffyBatch} class combines the information of various {\it CEL} files with a common {\it CDF} file. This class is designed to keep information of one experiment. The probe level data is contained in this object. The data in \verb+Dilution+ is a small sample of probe sets from 2 sets of duplicate arrays hybridized with different concentrations of the same RNA. This information is part of the \Robject{AffyBatch} and can be accessed with the \verb+phenoData+ and \verb+pData+ methods: <<>>= phenoData(Dilution) pData(Dilution) @ Several of the functions for plotting summarized probe level data are useful for diagnosing problems with the data. The plotting functions \Rfunction{boxplot} and \Rfunction{hist} have methods for \Robject{AffyBatch} objects. Each of these functions presents side-by-side graphical summaries of intensity information from each array. Important differences in the distribution of intensities are often evident in these plots. The function \Rfunction{MAplot} (applied, for example, to \verb+pm(Dilution)+), offers pairwise graphical comparison of intensity data. The option \verb+pairs+ permits you to chose between all pairwise comparisons (when \verb+TRUE+) or compared to a reference array (the default). These plots can be particularly useful in diagnosing problems in replicate sets of arrays. The function argument \verb+plot.method+ can be used to create a MAplot using a smoothScatter, rather than the default method which is to draw every point. \begin{figure}[htbp] \begin{center} <>= data(Dilution) MAplot(Dilution,pairs=TRUE,plot.method="smoothScatter") @ \end{center} \caption{Pairwise MA plots} \end{figure} \subsection{Accessing $PM$ and $MM$ Data} The $PM$ and $MM$ intensities and corresponding {\it affyID} can be accessed with the \Rfunction{pm}, \Rfunction{mm}, and \Rfunction{probeNames} methods. These will be matrices with rows representing probe pairs and columns representing arrays. The gene name associated with the probe pair in row $i$ can be found in the $i$th entry of the vector returned by \Rfunction{probeNames}. <<>>= Index <- c(1,2,3,100,1000,2000) ##6 arbitrary probe positions pm(Dilution)[Index,] mm(Dilution)[Index,] probeNames(Dilution)[Index] @ \verb+Index+ contains six arbitrary probe positions. Notice that the column names of $PM$ and $MM$ matrices are the sample names and the row names are the {\it affyID}, e.g. \verb+1001_at+ and \verb+1000_at+ together with the probe number (related to position in the target sequence). <<>>= sampleNames(Dilution) @ {\bf Quick example:} To see what percentage of the $MM$ are larger than the $PM$ simply type <<>>= mean(mm(Dilution)>pm(Dilution)) @ The \Rfunction{pm} and \Rfunction{mm} functions can be used to extract specific probe set intensities. <<>>= gn <- geneNames(Dilution) pm(Dilution, gn[100]) @ The method \Rfunction{geneNames} extracts the unique {\it affyID}s. Also notice that the 100th probe set is different from the 100th probe! The 100th probe is not part of the the 100th probe set. The methods \Rfunction{boxplot}, \Rfunction{hist}, and \Rfunction{image} are useful for quality control. Figure \ref{f3} shows kernel density estimates (rather than histograms) of $PM$ intensities for the 1st and 2nd array of the \verb+Dilution+ also included in the package. \subsection{Histograms, Images, and Boxplots} \begin{figure}[htbp] \begin{center} <>= hist(Dilution[,1:2]) ##PM histogram of arrays 1 and 2 @ \caption{\label{f3} Histogram of $PM$ intensities for 1st and 2nd array} \end{center} \end{figure} As seen in the previous example, the sub-setting method \verb+[+ can be used to extract specific arrays. {\bf NOTE: Sub-setting is different in this version. One can no longer subset by gene. We can only define subsets by one dimension: the columns, i.e. the arrays. Because the \verb+Cel+ class is no longer available \verb+[[+ is no longer available.} %]] The method \verb+image()+ can be used to detect spatial artifacts. By default we look at log transformed intensities. This can be changed through the \verb+transfo+ argument. <>= par(mfrow=c(2,2)) image(Dilution) @ \begin{figure}[htbp] \begin{center} \includegraphics{image} \caption{\label{f1} Image of the log intensities.} \end{center} \end{figure} These images are quite useful for quality control. We recommend examining these images as a first step in data exploration. The method \Rfunction{boxplot} can be used to show $PM$, $MM$ or both intensities. \begin{figure}[htbp] \begin{center} <>= par(mfrow=c(1,1)) boxplot(Dilution, col=c(2,3,4)) @ \caption{\label{f4}Boxplot of arrays in Dilution data.} \end{center} \end{figure} As discussed in the next section this plot shows that we need to normalize these arrays. \subsection{RNA degradation plots} The functions \Rfunction{AffyRNAdeg}, \Rfunction{summaryAffyRNAdeg}, and \Rfunction{plotAffyRNAdeg} aid in assessment of RNA quality. Individual probes in a probeset are ordered by location relative to the $5'$ end of the targeted RNA molecule.\cite{affy4} Since RNA degradation typically starts from the $5'$ end of the molecule, we would expect probe intensities to be systematically lowered at that end of a probeset when compared to the $3'$ end. On each chip, probe intensities are averaged by location in probeset, with the average taken over probesets. The function \Rfunction{plotAffyRNAdeg} produces a side-by-side plots of these means, making it easy to notice any $5'$ to $3'$ trend. The function \Rfunction{summaryAffyRNAdeg} produces a single summary statistic for each array in the batch, offering a convenient measure of the severity of degradation and significance level. For an example <<>>= deg <- AffyRNAdeg(Dilution) names(deg) @ does the degradation analysis and returns a list with various components. A summary can be obtained using <<>>= summaryAffyRNAdeg(deg) @ Finally a plot can be created using \Rfunction{plotAffyRNAdeg}, see Figure \ref{f4.3}. \begin{figure}[htbp] \begin{center} <>= plotAffyRNAdeg(deg) @ \caption{\label{f4.3} Side-by-side plot produced by plotAffyRNAdeg.} \end{center} \end{figure} \newpage \section{Normalization} \label{s1.4} Various researchers have pointed out the need for normalization of Affymetrix arrays. See for example \cite{bols:etal:2003}. The method \verb+normalize+ lets one normalize at the probe level <<>>= Dilution.normalized <- normalize(Dilution) @ For an extended example on normalization please refer to the vignette in the affydata package. \section{Classes} \label{classes} \verb+AffyBatch+ is the main class in this package. There are three other auxiliary classes that we also describe in this Section. \subsection{AffyBatch} The AffyBatch class has slots to keep all the probe level information for a batch of {\it Cel} files, which usually represent an experiment. It also stores phenotypic and MIAME information as does the \verb+ExpressionSet+ class in the Biobase package (the base package for BioConductor). In fact, \verb+AffyBatch+ extends \verb+ExpressionSet+. The expression matrix in \verb+AffyBatch+ has columns representing the intensities read from the different arrays. The rows represent the {\it cel} intensities for all position on the array. The cel intensity with physical coordinates\footnote{Note that in the {\it .CEL} files the indexing starts at zero while it starts at 1 in the package (as indexing starts at 1 in {\bf R}).} $(x,y)$ will be in row \[i = x + \mathtt{nrow} \times (y - 1)\]. The \verb+ncol+ and \verb+nrow+ slots contain the physical rows of the array. Notice that this is different from the dimensions of the expression matrix. The number of row of the expression matrix is equal to \verb+ncol+$\times$\verb+nrow+. We advice the use of the functions \verb+xy2indices+ and \verb+indices2xy+ to shuttle from X/Y coordinates to indices. For compatibility with previous versions the accessor method \verb+intensity+ exists for obtaining the expression matrix. The \verb+cdfName+ slot contains the necessary information for the package to find the locations of the probes for each probe set. See Section \ref{sec:probesloc} for more on this. \subsection{ProbeSet} The \verb+ProbeSet+ class holds the information of all the probes related to an {\it affyID}. The components are \verb+pm+ and \verb+mm+. The method \verb+probeset+ extracts probe sets from \verb+AffyBatch+ objects. It takes as arguments an \verb+AffyBatch+ object and a vector of {\it affyIDs} and returns a list of objects of class \verb+ProbeSet+ <<>>= gn <- featureNames(Dilution) ps <- probeset(Dilution, gn[1:2]) #this is what i should be using: ps show(ps[[1]]) @ The \verb+pm+ and \verb+mm+ methods can be used to extract these matrices (see below). This function is general in the way it defines a probe set. The default is to use the definition of a probe set given by Affymetrix in the CDF file. However, the user can define arbitrary probe sets. The argument \verb+locations+ lets the user decide the row numbers in the \verb+intensity+ that define a probe set. For example, if we are interested in redefining the \verb+AB000114_at+ and \verb+AB000115_at+ probe sets, we could do the following: First, define the locations of the $PM$ and $MM$ on the array of the \verb+1000_at+ and \verb+1001_at+ probe sets <<>>= mylocation <- list("1000_at"=cbind(pm=c(1,2,3),mm=c(4,5,6)), "1001_at"=cbind(pm=c(4,5,6),mm=c(1,2,3))) @ The first column of the matrix defines the location of the $PM$s and the second column the $MM$s. Now we are ready to extract the \verb+ProbSet+s using the \verb+probeset+ function: <<>>= ps <- probeset(Dilution, genenames=c("1000_at","1001_at"), locations=mylocation) @ Now, \verb+ps+ is list of \verb+ProbeSet+s. We can see the $PM$s and $MM$s of each component using the \verb+pm+ and \verb+mm+ accessor methods. <<>>= pm(ps[[1]]) mm(ps[[1]]) pm(ps[[2]]) mm(ps[[2]]) @ This can be useful in situations where the user wants to determine if leaving out certain probes improves performance at the expression level. It can also be useful to combine probes from different human chips, for example by considering only probes common to both arrays. Users can also define their own environment for probe set location mapping. More on this in Section \ref{sec:probesloc}. An example of a \verb+ProbeSet+ is included in the package. A spike-in data set is included in the package in the form of a list of \verb+ProbeSet+s. The help file describes the data set. Figure \ref{f5.3} uses this data set to demonstrate that the $MM$ also detect transcript signal. \begin{figure}[htbp] \begin{center} <>= data(SpikeIn) ##SpikeIn is a ProbeSets pms <- pm(SpikeIn) mms <- mm(SpikeIn) ##pms follow concentration par(mfrow=c(1,2)) concentrations <- matrix(as.numeric(sampleNames(SpikeIn)),20,12,byrow=TRUE) matplot(concentrations,pms,log="xy",main="PM",ylim=c(30,20000)) lines(concentrations[1,],apply(pms,2,mean),lwd=3) ##so do mms matplot(concentrations,mms,log="xy",main="MM",ylim=c(30,20000)) lines(concentrations[1,],apply(mms,2,mean),lwd=3) @ \caption{\label{f5.3}PM and MM intensities plotted against SpikeIn concentration} \end{center} \end{figure} \section{Location to ProbeSet Mapping} \label{sec:probesloc} On Affymetrix GeneChip arrays, several probes are used to represent genes in the form of probe sets. From a {\it CEL} file we get for each physical location, or cel, (defined by $x$ and $y$ coordinates) an intensity. The {\it CEL} file also contains the name of the {\it CDF} file needed for the location-probe-set mapping. The {\it CDF} files store the probe set related to each location on the array. The computation of a summary expression values from the probe intensities requires a fast way to map an {\it affyid} to corresponding probes. We store this mapping information in {\bf R} environments\footnote{Please refer to the {\bf R} documentation to know more about environments.}. They only contain a part of the information that can be found in the {\it CDF} files. The {\it cdfenvs} are sufficient to perform the numerical processing methods included in the package. For each {\it CDF} file there is package, available from \url{http://bioconductor.org/help/bioc-views/release/data/annotation/}, that contains exactly one of these environments. The {\it cdfenvs} we store the $x$ and $y$ coordinates as one number (see above). In instances of {\it AffyBatch}, the {\it cdfName} slot gives the name of the appropriate {\it CDF} file for arrays represented in the \verb+intensity+ slot. The functions \verb+read.celfile+, \verb+read.affybatch+, and \verb+ReadAffy+ extract the {\it CDF} filename from the {\it CEL} files being read. Each {\it CDF} file corresponds to exactly one environment. The function \verb+cleancdfname+ converts the Affymetrix given {\it CDF} name to a BioConductor environment and annotation name. Here are two examples: These give environment names: <<>>= cat("HG_U95Av2 is",cleancdfname("HG_U95Av2"),"\n") cat("HG-133A is",cleancdfname("HG-133A"),"\n") @ This gives annotation name: <<>>= cat("HG_U95Av2 is",cleancdfname("HG_U95Av2",addcdf=FALSE),"\n") @ An environment representing the corner of an Hu6800 array is available with the package. In the following, we load the environment, look at the names for the first 5 objects defined in the environment, and finally look at the first object in the environment: <<>>= data(cdfenv.example) ls(cdfenv.example)[1:5] get(ls(cdfenv.example)[1],cdfenv.example) @ The package needs to know what locations correspond to which probe sets. The \verb+cdfName+ slot contains the necessary information to find the environment with this location information. The method \verb+getCdfInfo+ takes as an argument an \verb+AffyBatch+ and returns the necessary environment. If \verb+x+ is an \verb+AffyBatch+, this function will look for an environment with name \verb+cleancdfname(x@cdfName)+. <<>>= print(Dilution@cdfName) myenv <- getCdfInfo(Dilution) ls(myenv)[1:5] @ By default we search for the environment first in the global environment, then in a package named \verb+cleancdfname(x@cdfName)+. Various methods exist to obtain locations of probes as demonstrated in the following examples: <<>>= Index <- pmindex(Dilution) names(Index)[1:2] Index[1:2] @ \verb+pmindex+ returns a list with probe set names as names and locations in the components. We can also get specific probe sets: <<>>= pmindex(Dilution, genenames=c("1000_at","1001_at")) @ The locations are ordered from 5' to 3' on the target transcript. The function \verb+mmindex+ performs in a similar way: <<>>= mmindex(Dilution, genenames=c("1000_at","1001_at")) @ They both use the method \verb+indexProbes+ <<>>= indexProbes(Dilution, which="pm")[1] indexProbes(Dilution, which="mm")[1] indexProbes(Dilution, which="both")[1] @ The \verb+which="both"+ options returns the location of the $PM$s followed by the $MM$s. \section{Configuring the package options} \label{configure.options} Package-wide options can be configured, as shown below through examples. \begin{itemize} \item Getting the names for the options: <<>>= opt <- getOption("BioC") affy.opt <- opt$affy print(names(affy.opt)) @ %$ \item Default processing methods: <<>>= opt <- getOption("BioC") affy.opt <- opt$affy affy.opt$normalize.method <- "constant" opt$affy <- affy.opt options(BioC=opt) @ %$ \item Compression of files: if you are always compressing your CEL files, you might find annoying to specify it each time you call a reading function. It can be specified once for all in the options. <<>>= opt <- getOption("BioC") affy.opt <- opt$affy affy.opt$compress.cel <- TRUE opt$affy <- affy.opt options(BioC=opt) @ %$ \item Priority rule for the use of a cdf environment: The option {\it probesloc} is a list. Each element of the list is itself a list with two elements {\it what} and {\it where}. When looking for the information related to the locations of the probes on the array, the elements in the list will be looked at sequentially. The first one leading to the information is used (an error message is returned if none permits to find the information). The element {\it what} can be one of {\it package}, {\it environment}. \end{itemize} \section{Where can I get more information?} \label{moreinfo} There are several other vignettes addressing more specialised topics related to the {\tt affy} package. \begin{itemize} \item {\bf affy: Custom Processing Methods (HowTo)}: A description of how to use custom preprocessing methods with the package. This document gives examples of how you might write your own preprocessing method and use it with the package. \item {\bf affy: Built-in Processing Methods}: A document giving fuller descriptions of each of the preprocessing methods that are available within the {\tt affy} package. \item {\bf affy: Import Methods (HowTo)}: A discussion of the data structures used and how you might import non standard data into the package. \item {\bf affy: Loading Affymetrix Data (HowTo)}: A quick guide to loading Affymetrix data into R. \item {\bf affy: Automatic downloading of cdfenvs (HowTo)}: How you can configure the automatic downloading of the appropriate {\it cdfenv} for your analysis. \end{itemize} \appendix \section{Previous Release Notes} \subsection{Changes in versions 1.6.x} There were very few changes. \begin{itemize} \item The function \verb+MAplot+ has been added. It works on instances of AffyBatch. You can decide if you want to make all pairwise MA plots or compare to a reference array using the pairs argument. \item Minor bugs fixed in the parsers. \item The path of celfiles is now removed by ReadAffy. \end{itemize} \subsection{Changes in versions 1.5.x} There are some minor differences in what you can do but little functionality has disappeared. Memory efficiency and speed have improved. \begin{itemize} \item The widgets used by ReadAffy have changed. \item The path of celfiles is now removed by ReadAffy. \end{itemize} \subsection{Changes in versions 1.4.x} There are some minor differences in what you can do but little functionality has disappeared. Memory efficiency and speed have improved. \begin{itemize} \item For instances of \verb+AffyBatch+ the subsetting has changed. For consistency with \verb+exprSets+ one can only subset by the second dimension. So to obtain the first array, \verb+abatch[1]+ and \verb+abatch[1,]+ will give warnings (errors in the next release). The correct code is \verb+abatch[,1]+. \item mas5calls is now faster and reproduces Affymetrix's official version much better. \item If you use \verb+pm+ and \verb+mm+ to get the entire set of probes, e.g. by typing \verb+pm(abatch)+ then the method will be, on average, about 2-3 times faster than in version 1.3. \end{itemize} \bibliographystyle{plainnat} \bibliography{affy} \end{document} affy/inst/doc/affy.pdf0000644000175200017520000301631014516022624015671 0ustar00biocbuildbiocbuild%PDF-1.5 % 128 0 obj << /Length 1103 /Filter /FlateDecode >> stream xXKoFW%`n%s),I]Hib ȡa+%")TwX -vٙoޜ,.^<>BS!uʜ,E+ye[nD48ߘ_?qEJp)7<zA . 4f:#4ႍ~P(cp =@S'Jȏ0ȀɬDH0ɂ|3Az '7cѬ‡Ow\,;ȟd8VnkCC D/8gpv&IHnqW?lU>zb(r*^MYkHڸ 71jڀfwW^ ˥ސP$yx԰ :\<T)<Nz>ANjs S(,$|nfMwP^~&הW:Éje&d+p yqF:tHOf'蔇qBcxk й%tOZ4.u‹` C1@ᒩVkG{_? }︿:x? =,\v*='X&> ԾZ}ݯ]ҧCW \gZ6\[ M39h~D`T$*pzq7qn endstream endobj 158 0 obj << /Length 2265 /Filter /FlateDecode >> stream xYKo6W-j)Q<3ɮ ;1$YbwsWD_*K.nY*>|;Y.wi.RЯv?7C^=n(UbP3 E$Bl?>|-)Rh!9_4GW2PQ#1>eyF6a> wTci.[a6 gE`vJ,#XlQuXp()K'E^;&{CH欨Qwz'MKY2G&Y,]̢hR{C%<f2Jƍ#:!Mv/Pt4dNүz1C-^",˖|N|@ x}6;#Ak*LACE[ԯ*wc ` lhh;k\sc}(j}ꬮc;j]+@_(`ƙ=RIz4b3{Eh< vCw4qYY+Uڜehg6Si^1{T+I P@DA jufPqȬ}3 5 ,4H14GØDS,xREwI %s e,f"K!v)Hb;!,>l[MH\N(A<M.r1(P6< >M ;h$>ϲNJ#G X ;\:IgSߍ~^6¢RU@ 6!ŀxRhbhap6P]6~mnҥ7Pʱ?dN[:fYd^׼aIH^RF(I6h{]H"3>}#-t뢧)~n~BfO־qf:o-} < k # zٹ8;-rwC [P2EOy<"Q76:(; ض[j}c.EOSTฯ0v: (ӟZw @xK ?WƪAQrd[hܺ^QnĎb GΠUlK8 \5SG_@?ҵ0eikWo-E0 Z> stream xZms6_otH,ޑK$sӻKAGS[Hr߳4))-v]B$!Z 'H4F^k+( P$DQ(\W! r(hK ׵FU,CY0:50J^ =E+WE*)[L1=@VG8H gk}PA * Q,BjIEaI D(aKs.%K7EdU€$/-l# lk 4Ih$ԷTI|% Kc`20A+ -b CJ/4VКeA %R@#ETmlL;2y9І7  "; ) RiV5I`_ƃJ ==0s=2^DV  #0'2Ȃ<`b>< eAuČLU-`e& 08:(>T|&ǣŤX'>7Sn̸9憛Q;w;'Qۓ<+7<7cn|Fr%O1[]}C]O2zz$P|;A"-^=QﳉRDvB%n$<~(g)Xp^%zNvBr2Y xw#f@`(:j?`csNYXX‰cs 15 Th , &_Q;-h6Iwjt:P2={!fYKZʺ?ZKTTTTTTۘT+IϙT+T+$&96ɱVIMRDjTJ#JxVm. &:`o<8M฽D8ܡcRe$]kD'㬰Q n_j6XM(U/4YN:h _@F`<^mW%>}*HX"xa RmJW55 (#+QK8 2<\E20C3M~2^ /ɹ2+@5 )*Af|]*.Qskɛ"eZcsCd\<~51rSk}W 4AFgM Q:#(OşG_'eyRhQ͊jvQ>nZN4F2P5(@, endstream endobj 185 0 obj << /Length 2308 /Filter /FlateDecode >> stream xڕXKܶWlTy0$C\l9NE9[C,qHfӍ Q.Ct75~s ]RxeEΊ??m>,Ib|,ceϣQn/<:>0ȺqM -1~hbޯvfET5@ZylH =.yuOqQ;X`H`Bd$mw]3tm=e ~iq6ߦRRJ-Zq`֬`?lytVAКrXh|9zr#FK+`2/ItEdS롲g ؀'R9[Q6ϻpi&'7|\$ ش~ϱ'$9\{¬/(Oy.)Z: M /D0Gb#]NQQDa9[= ^ B ( s9^KoBHG\^Mm0uR];P¥t1Ν\ >7 ;:m{Y [3ǥ"Qfo\r.gqRjթa $jU_vRDQV3U~|f'u=sFD\4i-7x%>~#;4yqq _-1E?z6,zěG5uOgG*g6rcCEHz?e\nt¯]}n[t֙lͨnLȹ &)Di4Fglk֏v_&:FRgOִ5P*u4N>M@"g9WRӨ!:kK^)$V jM?&i?eE2"UOo}|$F%l6?iU;_db9#7_fAKqhhn )mյG|?A@G me3|M?{:GD\BDZӚxH'WjuD"9k\mSfif^,9VI=t-(o2Ek>} 8Y. -K9U"% 뫒 1SVD~*D܂&k_3h~_5l6- [ +"vhgkvI<xU/ˑߡOd%^3~ث q7 6w7Om 1olYMɅs;.80NF INJ\vfA辶N>_\ V00B@=[8GDkx_`:jT4r6QB_`a` !BC B^c%a \/&:ӵ|LsGG-}*ɥ}o`BېY̎/'R 6.?ghuU#|)}ILN"$" @/OtE͕~`QV J}z5eҧ.@: vdd?]|OP=&r ghg).3 CGn>n夔S~QPd/h<'8:boeU ȶ`3緇Ck*Sxhtwv=uy8~wZjftMabĩ++qhם:ʵ!|'g7um dû|}+j-x)KY.O7y%I4F/cغ -΄2 _,7^| &m(R蝌Azп{zm[Y"Ǧ`AwGF/:-@FiHȸ\?C2ͣme`i="L,儌z8,n]di6{O N\3;M6UCQ6L8i4pfAɲ,[U"(=q4s( `o LO endstream endobj 199 0 obj << /Length 2154 /Filter /FlateDecode >> stream xڕXY~_a`VQ))$؞`ge۴T:Lb*͛Znd"DG&EY*ևA,v'e? ~yuti/}>7R\)wGEZ@/%M^Q4-^Q{܉/}CcTDqiNXRyLoh9T!I$b-,IprE(I*=0NJr._B˥Jv]Z mCJE7;@hb5E: axAx;/STyd;Ҍ)ܰPH0t31z>Q(`x6)-AB50 vJ4QK$s~O! 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It returns the object unchanged. May be used as a placeholder. \subsection{rma/rma2} These are background adjustment implementations for the rma method \cite{iriz:etal:2003}. They differ only in how they estimate a set of parameters (generally you should use \verb+rma+ in preference to \verb+rma2+. In both cases PM probe intensities are corrected by using a global model for the distribution of probe intensities. The model is suggested by looking at plots of the empirical distribution of probe intensities. In particular the observed PM probes are modeled as the sum of a normal noise component N (Normal with mean $\mu$ and variance $\sigma^2$) and a exponential signal component S (exponential with mean $\alpha$). To avoid any possibility of negatives, the normal is truncated at zero. Given we have O the observed intensity, this then leads to an adjustment. \begin{equation*} E\left(s \lvert O=o\right) = a + b \frac{\phi\left(\frac{a}{b}\right) - \phi\left(\frac{o-a}{b}\right)}{\Phi\left(\frac{a}{b}\right) + \Phi\left(\frac{o-a}{b}\right) - 1 } \end{equation*} where $a = s- \mu - \sigma^2\alpha$ and $b = \sigma$. Note that $\phi$ and $\Phi$ are the standard normal distribution density and distribution functions respectively. Note that MM probe intensities are not corrected by either of these routines. \subsection{mas} This is an implementation of the background correction method outlined in the Statistical Algorithms Description Document \cite{affy:tech:2002}. The chip is broken into a grid of 16 rectangular regions. For each region the lowest 2\% of probe intensities are used to compute a background value for that grid. Each probe is then adjusted based upon a weighted average of the backgrounds for each of the regions. The weights are based on the distances between the location of the probe and the centriods of 16 different regions. Note this method corrects both PM and MM probes. \section{Normalization Methods} You can see the background correction methods that are built into the package by examining the variable \verb+bgcorrect.method+. <<>>= normalize.AffyBatch.methods() @ The Quantile, Contrast and Loess normalizations have been discussed and compared in \cite{bols:etal:2003}. \subsection{quantiles/quantiles.robust} The quantile method was introduced by \cite{bols:etal:2003}. The goal is to give each chip the same empirical distribution. To do this we use the following algorithm where $X$ is a matrix of probe intensities (probes by arrays): \begin{enumerate} \item Given $n$ array of length $p$, form $X$ of dimension $p \times n$ where each array is a column \item Sort each column of $X$ to give $X_{\mbox{sort}}$ \item Take the means across rows of $X_{\mbox{sort}}$ and assign this mean to each element in the row to get $X'_{\mbox{sort}}$ \item Get $X_{\mbox{normalized}}$ by rearranging each column of $X'_{\mbox{sort}}$ to have the same ordering as original $X$ \end{enumerate} The quantile normalization method is a specific case of the transformation $x'_{i} = F^{-1}\left(G\left(x_{i}\right)\right)$, where we estimate $G$ by the empirical distribution of each array and $F$ using the empirical distribution of the averaged sample quantiles. Quantile normalization is pretty fast. The {\tt quantiles} function performs the algorithm as above. The {\tt quantile.robust} function allows you to exclude or down-weight arrays in the computation of $\hat G$ above. In most cases we have found that the {\tt quantiles} method is sufficient for use and {\tt quantiles.robust} not required. \subsection{loess} There is a discussion of this method in \cite{bols:etal:2003}. It generalizes the $M$ vs $A$ methodology proposed in \cite{Dudoit:2002} to multiple arrays. It works in a pairwise manner and is thus slow when used with a large number of arrays. \subsection{contrasts} This method was proposed by \cite{astr:2003}. It is also a variation on the $M$ vs $A$ methodology, but the normalization is done by transforming the data to a set of contrasts, then normalizing. \subsection{constant} A scaling normalization. This means that all the arrays are scaled so that they have the same mean value. This would be typical of the approach taken by Affymetrix. However, the Affymetrix normalization is usually done after summarization (you can investigate \verb+affy.scalevalue.exprSet+ if you are interested) and this normalization is carried out before summarization. \subsection{invariantset} A normalization similar to that used in the dChip software \cite{li:wong:2001a}. Using a baseline array, arrays are normalized by selecting invariant sets of genes (or probes) then using them to fit a non-linear relationship between the ``treatment'' and ``baseline'' arrays. The non-linear relationship is used to carry out the normalization. \subsection{qspline} This method is documented in \cite{workman:etal:2002}. Using a target array (either one of the arrays or a synthetic target), arrays are normalized by fitting splines to the quantiles, then using the splines to perform the normalization. \section{PM correct methods} <<>>= pmcorrect.methods() @ \subsection{mas} An {\it ideal mismatch} is subtracted from PM. The ideal mismatch is documented by \cite{affy:tech:2002}. It has been designed so that you subtract MM when possible (ie MM is less than PM) or something else when it is not possible. The Ideal Mismatch will always be less than the corresponding PM and thus we can safely subtract it without risk of negative values. \subsection{pmonly} Make no adjustment to the pm values. \subsection{subtractmm} Subtract MM from PM. This would be the approach taken in MAS 4 \cite{affy4}. It could also be used in conjunction with the Li-Wong model. \section{Summarization methods} <<>>= express.summary.stat.methods() @ \subsection{avgdiff} Compute the average. This is the approach that was taken in \cite{affy4}. \subsection{liwong} This is an implementation of the methods proposed in \cite{li:wong:2001a} and \cite{li:wong:2001b}. The Li-Wong MBEI is based upon fitting the following multi-chip model to each probeset \begin{equation} y_{ij} = \phi_i \theta_j + \epsilon_{ij} \end{equation} where $y_{ij}$ is $PM_{ij}$ or the difference between $PM_{ij}-MM_{ij}$. The $\phi_i$ parameter is a probe response parameter and $\theta_j$ is the expression on array $j$. \subsection{mas} As documented in \cite{affy:tech:2002}, a robust average using 1-step Tukey biweight on $\log_2$ scale. \subsection{medianpolish} This is the summarization used in the RMA expression summary \cite{iriz:etal:2003}. A multichip linear model is fit to data from each probeset. In particular for a probeset $k$ with $i=1,\dots,I_k$ probes and data from $j=1,\dots,J$ arrays we fit the following model \begin{equation*} \log_2\left(PM^{(k)}_{ij}\right) = \alpha_i^{(k)} + \beta_j^{(k)} + \epsilon_{ij}^{(k)} \end{equation*} where $\alpha_i$ is a probe effect and $\beta_j$ is the $\log_2$ expression value. The medianpolish is an algorithm (see \cite{tukey:1977}) for fitting this model robustly. Please note that expression values you get using this summary measure will be in $\log_2$ scale. \subsection{playerout} This method is detailed in \cite{Lazardis:etal:2002}. A non-parametric method is used to determine weights. The expression value is then the weighted average. \section{Putting it altogether using {\tt expresso}} The function that you should use is {\tt expresso}. It is important to note that not every preprocessing method can be combined together. In particular the \verb+rma+ backgrounds adjust only PM probe intensities and so they should only be used in conjunction with the \verb+pmonly+ PM correction. Also remember that the \verb+mas+ and \verb+medianpolish+ summarization methods $\log_2$ transform the data, thus they should not be used in conjunction with any preprocessing steps that are likely to yield negatives like the \verb+subtractmm+ pm correction method. The following is a typical call to \verb+expresso+. \begin{Sinput} library(affydata) data(Dilution) eset <- expresso(Dilution,bgcorrect.method="rma", normalize.method="quantiles", pmcorrect.method="pmonly", summary.method="medianpolish") \end{Sinput} %@ This would give you the RMA expression measure, but of course there are other ways of computing RMA (chiefly \verb+rma+). The true power of \verb+expresso+ becomes apparent when you start combining different methods. By choosing a method for each of the four steps ({\tt bgcorrect.method}, {\tt normalize.method}, {\tt pmcorrect.method}, {\tt summary.method}) you can create quite a variety of expression measures. For instance \begin{Sinput} eset <- expresso(Dilution,bgcorrect.method="mas", normalize.method="qspline", pmcorrect.method="subtractmm", summary.method="playerout") \end{Sinput} would be a valid way of computing an expression measure (it is up to the user to decide whether such a concoction is sensible or not). \bibliographystyle{plainnat} \bibliography{affy} \end{document} affy/inst/doc/builtinMethods.pdf0000644000175200017520000056110114516022626017740 0ustar00biocbuildbiocbuild%PDF-1.5 % 120 0 obj << /Length 812 /Filter /FlateDecode >> stream xX]o0}߯ȣ#_Ix, $ڳ g\hx-H&i0_H q4̺ji ` 9mLH&yamZ?O+ Ad,*I|Y\BLfmpuV(T [sֻ/g.x[θ"%]D 1 Gcgf F?T% 0SeZ#"ܛ@HSKҺsm(OR̽!!g gY,?-ӭh+R{#>u}$r(zLԃSƀ̪NBFZlc̚:;/uGQY >#{y`Uo'۴ZʹUcҖUa(GQc:ۆ4Q%+TlH\9 PHc`{qBNȍ΍/&~T2@eBpac!C#ZhKɚrn+_XqtP'vXījnZ+7MhumVՅ;R#\ U8o > stream xڵYKsW X1ǘ^*rjIKjw 9" 2,|==*N a滿l%ddVw$jHCۯra?Vk]S7xϻǻDRdQDڨXԟ;-HqT|*v[KQ}+ng۶ ?p$xu׀!sНIЕOL؞S0s5FH#Yˉ QG"LĐ7diI1*J{|G }^GQC(~pUi=(K^ʼ)xs_7G{CN5GC+*2pؘD$rml[~hC{ cvG}<`0.vyu}ͩ8%=zwLH0bO0E9}h/8Ȯ>mQ莌tF8I`1ETjܞ[Q]]y1&%$|_tX(=_K)CF k.~/碱{H fכHFAYlyF!E^oHVFrA$*3o%0 5V3ȟdgd<8٤X1 uvW,o'AUcvj{Җ5ԇl B3\qfͫ#6/ivk0'2c]ԖjS^mwSP*KiqS,sۍ:CiO W+ pÐ& ?6oy<1bIU* 59) @g(BuUzKՀ ?f`8Ic3*xM{`+xP.c}.W̽_JҥZ=e^@jNe/:jU6IR T|i >[z"e6F1gV[EWȜg bHy>V4 R䰲%N>ڭ{?Z@Eۡ9ng`<NfCQ-&^ޞ8] 1 J ?\v:ugXC;':3C _*VּMhzzچ[}X:#V?{6OL;11xlɠ.ZOY;>`Dkks?QIP?*e8!_\Oq@ϗ.U /g*{VFD.5.Oh6>.U2ȑ\5TqJXSctfzzS:>xJ HPJ_fw. 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Custom Processing Methods} %\VignetteKeywords{Preprocessing, Affymetrix} %\VignetteDepends{affy} %\VignettePackage{affy} %documentclass[12pt, a4paper]{article} \documentclass[12pt]{article} \usepackage{amsmath} \usepackage{hyperref} \usepackage[authoryear,round]{natbib} \textwidth=6.2in \textheight=8.5in %\parskip=.3cm \oddsidemargin=.1in \evensidemargin=.1in \headheight=-.3in \newcommand{\scscst}{\scriptscriptstyle} \newcommand{\scst}{\scriptstyle} \newcommand{\Rfunction}[1]{{\texttt{#1}}} \newcommand{\Robject}[1]{{\texttt{#1}}} \newcommand{\Rpackage}[1]{{\textit{#1}}} \newcommand{\Rmethod}[1]{{\texttt{#1}}} \newcommand{\Rfunarg}[1]{{\texttt{#1}}} \newcommand{\Rclass}[1]{{\textit{#1}}} \author{Laurent} \begin{document} \title{affy: Custom Processing Methods (HowTo)} \maketitle \tableofcontents \section{Introduction} This document describes briefly how to customize the affy package by adding one's own processing methods. The types of processing methods are background correction, normalization, perfect match correction and summary expression value computation. We tried our best to make this as easy as we could, but we are aware that it is far from being perfect. We are still working on things to improve them. Hopefully this document should let you extend the package with supplementary processing methods easily. As usual, loading the package in your \verb+R+ session is required. \begin{Sinput} R> library(affy) ##load the affy package \end{Sinput} <>= library(affy) @ \section{How-to} For each processing step, labels for the methods known to the package are stored in variables. <<>>= normalize.AffyBatch.methods() bgcorrect.methods() pmcorrect.methods() express.summary.stat.methods() @ We would recommend the use of the method \verb+normalize.methods+ to access the list of available normalization methods (as a scheme for normalization methods that would go beyond 'affy' is thought). <<>>= library(affydata) data(Dilution) normalize.methods(Dilution) @ For each processing step, a naming convention exists between the method label and the function name in \verb+R+ (see table~\ref{table:summary.labels}). Each processing methods should be passed objects (and return objects) corresponding to the processing step (see table~\ref{table:summary.methods}). \begin{table} \begin{tabular}{|c|c|} \hline variable for labels & naming convention \\ \hline bgcorrect.methods & bg.correct.