Yeah, they just factorized the colData or whatever the equivalent is in the older version that you're using (I'll have to download that version and have a look).
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So the first thing to try is to just reset ecs@designColumns to contain factors (if they don't already). This won't solve the original issue, since I suspect that the conversion to a dataframe is occurring after this step. The quickest way to go about this is to just look at the code for estimatelog2FoldChanges (I'll paste it below) and just execute every line manually. Then you'll know exactly what it's processing and why the duplicate row name error is happening (this will also give you the chance to avert that error then).
Code:estimatelog2FoldChanges <- function(ecs, fitExpToVar="condition", denominator="", getOnlyEffects=FALSE, averageOutExpression=TRUE, nCores=1, quiet=FALSE, file="") { stopifnot(is(ecs, "ExonCountSet")) if(any(is.na(sizeFactors(ecs)))){ stop("Please estimate sizeFactors first\n")} if(!fitExpToVar %in% ecs@designColumns){ stop("fitExpToVar parameter is not in the design columns, double check ecs@designColumns")} if(sum(is.na(featureData(ecs)$dispersion))==nrow(counts(ecs))){ stop("No dispersion parameters found, first call function estimateDispersions...\n")} frm <- as.formula(paste("count ~", fitExpToVar, "* exon")) testablegenes <- as.character(unique(fData(ecs)[which(fData(ecs)$testable),]$geneID)) geteffects <- function(geneID){ coefficients <- fitAndArrangeCoefs(ecs, geneID=geneID, frm, balanceExons=TRUE) if( is.null( coefficients ) ){ return(coefficients) } ret <- t(getEffectsForPlotting(coefficients, averageOutExpression=averageOutExpression, groupingVar=fitExpToVar)) rownames(ret) <- paste(geneID, rownames(ret), sep=":") return(ret) } if( nCores > 1 ){ if(!is.loaded("mc_fork", PACKAGE="parallel")){ stop("Please load first parallel package or set parameter nCores to 1...")} alleffects <- parallel:::mclapply( testablegenes, function(x){ geteffects(x) }, mc.cores=nCores ) }else{ alleffects <- lapply( testablegenes, function(x){geteffects(x)}) } names(alleffects) <- testablegenes alleffects <- do.call(rbind, alleffects) alleffects <- vst(exp( alleffects ), ecs) toadd <- matrix(NA, nrow=nrow(ecs), ncol=ncol(alleffects)) rownames(toadd) <- featureNames(ecs) if( getOnlyEffects ){ colnames(toadd) <- colnames(alleffects) toadd[rownames(alleffects), colnames(alleffects)] <- alleffects }else{ if( denominator == "" ){ denominator <- as.character(design(ecs, drop=FALSE)[[fitExpToVar]][1]) } stopifnot( any( colnames(alleffects) %in% denominator ) ) denoCol <- which(colnames(alleffects) == denominator) alleffects <- log2(alleffects / alleffects[,denoCol]) colnames(alleffects) <- sprintf("log2fold(%s/%s)", colnames(alleffects), denominator) colnames(toadd) <- colnames(alleffects) alleffects <- alleffects[,-denoCol, drop=FALSE] toadd <- toadd[,-denoCol, drop=FALSE] toadd[rownames(alleffects), colnames(alleffects)] <- alleffects } fData(ecs) <- cbind(fData(ecs), toadd) ecs }
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by GATTACATLove this - good data definitely starts from good input, and poor input can only give relatively poor data. I particularly like the mention of Nanodrop/absorbance based methods for quantification. It's such a toss up if you'll get an accurate reading or what amounts to a randomly generated number, and a lot of library/sequencing related issues can be traced back to poor quant.
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07-01-2026, 11:43 AM -
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by SEQadmin2
I’m not a sequencing expert. I’m a purification scientist who uses NGS to evaluate workflows my group develops. With this perspective, we think about the sample first and the NGS workflow second. The sequencer is an exceptionally honest reporter, but it can only report on what you give it, so whether you get clean, interpretable data from an NGS workflow is largely determined before you begin.
Here are nine questions we think about, in roughly the order they matter, before...-
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