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  • How to deal with samples partially without replicates in DESeq?

    I have four groups of samples, 2 groups with replicates and 2 groups without. The data frame is like this:
    expt_design<-data.frame(row.name=colnames(colon)
    + condition=c("GF","SPF","9343","9343","SPT","SPT"))
    Error: unexpected symbol in:
    "expt_design<-data.frame(row.name=colnames(colon)
    condition"
    > expt_design<-data.frame(row.names=colnames(colon),
    + condition=c("GF","SPF","9343","9343","SPT","SPT"))
    > expt_design
    condition
    GF GF
    SPF SPF
    X9343.1 9343
    X9343.2 9343
    SPT.1 SPT
    SPT.2 SPT
    > conditions=expt_design$condition
    > conditions
    [1] GF SPF 9343 9343 SPT SPT
    Levels: 9343 GF SPF SPT

    When I tried to run it gave me this error message:
    >cds=estimateDispersions(cds)
    Error in .local(object, ...) :
    None of your conditions is replicated. Use method='blind' to estimate across conditions, or 'pooled-CR', if you have crossed factors.

    Plz help me to identify the problem and tell me how to deal with partially replicated samples

  • #2
    Did you read the error message, the fix is contained within it. Also, you should read the DESeq tutorial.

    Comment


    • #3
      You might have confused DESeq by passing your "condition" data as a data.frame rather than as a factor. Try again using

      cds <- newCountDataSet( countTable, exptDesign$condition)

      And consider switching to DESeq2. It not only has improved power; we have also improved the interface to avoid problems like this one.

      Comment

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