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  • Name_less_
    Member
    • Oct 2014
    • 12

    #1

    Help with DESeq please

    Hello everyone
    I am new in the use of DESeq and I have problems for obtain Diferencial Expression of an Output of EDGE-Pro.
    In R, I followed the steps on this page:

    https://wikis.utexas.edu/display/bio...ssion+analysis

    In where are the next steps:

    login1$ R
    ...
    > library("DESeq")
    > counts = read.delim("gene_counts.tab", header=F, row.names=1)
    > head(counts)
    > colnames(counts) = c("wt1", "mut1", "wt2", "mut2")
    > head(counts)
    > my.design <- data.frame(row.names = colnames( counts ),condition = c( "wt", "mut", "wt", "mut"), libType = c( "single-end", "single-end", "single-end", "single-end" ))
    > conds <- factor(my.design$condition)
    > cds <- newCountDataSet( counts, conds )

    BUT, in this point, I have the next error:

    Error en round(countData) : non-numeric argument to mathematical function

    Someone can help me? please

    My file is the next (the firsts five lines):

    gene 571A.rpkm_0 571B.rpkm_0 571C.rpkm_0 571D.rpkm_0
    geneA 406 609 423 477
    geneB 67 117 76 64
    geneC 21134 18099 14720 14153
    geneD 810 857 725 691

    Each 571 is one condition, without replicates.
  • edm1
    Member
    • May 2013
    • 10

    #2
    Is counts formatted correctly, does
    Code:
    head(counts)
    look like you'd expect?

    Is there a reason you are not using DESeq2? In DESeq2 you'd use DESeqDataSetFromMatrix instead of newCountDataSet:

    Code:
    dds <- DESeqDataSetFromMatrix(countData = counts,
                                  colData = my.design,
                                  design = <formula>)
    You should follow the PDFs available here:
    Estimate variance-mean dependence in count data from high-throughput sequencing assays and test for differential expression based on a model using the negative binomial distribution.

    Comment

    • Name_less_
      Member
      • Oct 2014
      • 12

      #3
      The table looks as expected, but I will try of use DESeq2. However, still I can not solve this problem in DESEQ.

      Comment

      • evozoa
        Junior Member
        • May 2014
        • 1

        #4
        Check to make sure your sample and gene names do not include spaces. If they do, it will throw off the column numbers.

        Comment

        • Name_less_
          Member
          • Oct 2014
          • 12

          #5
          Hi
          Finally I used DEseq2 and I could get what I expected. Thanks for all and greetings !!

          Comment

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