DESeqDataSetFromMatrix
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Gian ▴ 30
@chloe-25118
Last seen 2.3 years ago
Italy
control = factor(c(rep("Control",5),NA,NA))
affected=  factor(c(rep("Affected",7)))
library(DESeq2)
dds<-DESeqDataSetFromMatrix(
    countData=countTable,
    design =~control+affected,
    colData=data.frame(
      control=control,
      affected=affected
    ))
normCounts<-rlog(dds,blind=false)

This error coming

Error in DESeqDataSetFromMatrix(countData = countTable, design = ~control +  : 
  ncol(countData) == nrow(colData) is not TRUE
DESeqDataSetFromMatrix • 18k views
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2
Entering edit mode
Kevin Blighe ★ 4.0k
@kevin
Last seen 16 days ago
Republic of Ireland

Hi,

As implied by the error message, for this command to work correctly, the following must return TRUE:

meta <- data.frame(
  control = control,
  affected = affected)
ncol(countTable) == nrow(meta)

That is, to say, the number of columns of your input raw counts table (passed to the function as countData) must be equal to the number of rows of your input sample metadata (passed to the function as colData). It also follows that the order of these (the columns and the rows) must match.

As a suggestion, I think that you may need:

dds <- DESeqDataSetFromMatrix(
  countData = countTable,
  design = ~ condition,
  colData = data.frame(
    condition = c(control, affected)))

, assuming that the elements of c(control, affected) are perfectly aligned to the columns of countTable

Kevin

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0
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#normalizing counts
library(DESeq2)
dds<-DESeqDataSetFromMatrix(
    countData=countTable,
    design =~ condition,
    colData=data.frame(
      condition = c(control, affected)
    ))
normCounts<-rlog(dds,blind = FALSE)

When I add like this it shows this error for design Now I check that number of columns of countData is equal to the number of rows of colData

Error in DESeqDataSet(se, design = design, ignoreRank) : 
  design has a single variable, with all samples having the same value.
  use instead a design of '~ 1'. estimateSizeFactors, rlog and the VST can then be used
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2
Entering edit mode

Hi again, and thank you for replying.

control = factor(c(rep("Control",5),NA,NA))
affected = factor(c(rep("Affected",7)))

Why are there values of NA here? If you have samples whose condition is unknown, you may consider removing these samples from the dataset, or create a third 'condition' called 'Unknown'.



You may wish to try:

control = c(rep('Control', 5), 'Unknown', 'Unknown')
affected = rep('Affected', 7)
meta <- data.frame(
  condition = factor(c(control, affected),
    levels = c('Control', 'Affected', 'Unknown')))

dds <- DESeqDataSetFromMatrix(
  countData = countTable,
  design = ~ condition,
  colData = meta)
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1
Entering edit mode

Thanks, it resolved.

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