Question: export edgeR normalised OTU table
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Pozdrawiam • 0
@Pozdrawiam-24676
Last seen 22 days ago

Hello, I want to normalise my microbiome data (OTU table) using edgeR package and export my normalised matrix ( I want to test some normalisation methods, normalised data will serve to calculate beta diversity and visualise by PCoA). I went through bioconductor forum and biostars and I still don't know want is the proper way to export normalised data from edgeR.

So, I read my matrix and calcNorm:

d <- DGEList(counts = d, group=group)
d = calcNormFactors(d)

what happens next?:

1st option

d = estimateCommonDisp(d, verbose=TRUE)
normalised_df <- d$pseudo.alt

2nd option

normalised_df <- cpm(d,  normalized.lib.sizes = T)

3rd option

dgList <- estimateCommonDisp(dgList)
dgList <- estimateTagwiseDisp(dgList)
normalised_df <- t(t(dgList$pseudo.counts)*(dgList$samples$norm.factors))

I have an impression that every post claims something different and at this point I am very confused. Thank you!

microbiome edgeR • 99 views
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Is anything wrong with the answers you were provided here (https://www.biostars.org/p/487419/) citing the manual that 2nd option is preferred and third is discouraged? It is good practice to indicate crossposts by adding links to the other community.

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I just wanted to get a 3rd opinion, as your answer was different from the first one:)

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