can the counts extracted from Diffbind be used to analysis in Deseq2 as the RNAseq?
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@qq809825706-21501
Last seen 10 months ago
China

the consensus peak set extracted from diffbind is the counts of peak. if using the DESEQ method, diffbind will make analysis. so, can I directly use Deseq2 to analyze the consensus peak set. there are more other packages can analysis with Deseq2.

deseq2 diffbind • 1.7k views
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Rory Stark ★ 5.2k
@rory-stark-5741
Last seen 4 weeks ago
Cambridge, UK

You can extract the full count matrix and use it in other packages:

myDBA <- dba.count(myDBA, score=DBASCOREREADS) counts <- dba,peakset(myDBA, bRetrieve=TRUE)

In the upcoming release, to can also easily extract the DESeq2 analysis object itself.

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is there difference between the results made by Deseq2(with full count matrix ) and Diffbind(DBA_DESEQ2)?

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The default mode in DiffBind is to do a DESeq2 analysis, so it is the "same" in that sense. However there are many ways to do an analysis with DESeq2, using different designs, contrasts, normalization, options to estimate dispersion, fit a model, perform difference tests, etc. So you can get one specific type of DESeq2 analysis out of DiffBind, but using the reads counts to do a DESeq2 analysis yourself allows you to have complete control over these choices.

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OK, I see. Thank you very much.

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Hello, I am having a bit of trouble doing the contrasts with diffbind (I did post another question on this so hope this is nor cross posting), so it might be easier for me to export the count matrix made on the consensus peak to Deseq2. I have my count matrix like this : K4_count <- dba.count(K4, bUseSummarizeOverlaps=TRUE, bRemoveDuplicates = TRUE, bParallel = TRUE). So if I understand correctly, I just need to make counts = dba.peakset(K4, bRetrieve=TRUE) ? And this matrix I can feed the DESEQ2 tool right? Sorry if I'm being obtuse, I'm not confortable yet with R or coding in any way (still learning).

Thank you

Rita

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This will work, but you should also set the score parameter in dba.count() to score = DBA_SCORE_READS to get non-normalized read counts. The call to dba.peakset() will return a GRanges object, with the counts stored in the mcols() attribute.

You can also get a fully-formed DESeq2 DESeqDataSet object back using dba.analyze() with bRetrieveAnalysis=TRUE after doing a simple analysis, then use that to expand the design/contrast options directly in DESeq2.

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Thank you!

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