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  • gtosser
    Junior Member
    • Sep 2014
    • 4

    #1

    Use of glmer.nb to estimate mixed model effects on RNAseq data

    Hello,

    This is my first post on this site but I have already read many interesting posts here and I wish to thank all the participants.

    I have a RNAseq dataset generated with different factors:
    - species: 2 levels (pig, cattle)
    - treatment: 2 levels (control, treated)
    - animal: 10 levels
    I want to know what are the differentially expressed genes between control and treated conditions and if there is a species effect and also if there are species*gene interactions
    I have normalized the data (11882 genes) with Deseq2 and extracted the data from Deseq2

    >dds<-estimateSizeFactors(dds)
    >dds<-estimateDispersions(dds)
    >dds<-nbinomWaldTest(dds)
    >normalizedCounts <- t( t(counts(dds)) / sizeFactors(dds) )
    >dim(normalizedCounts)
    # [1] 11882 40

    After that I have constructed data1 where all the normalizedCounts are in column2 and the factors in columns 3,4,5. Column 1 is the gene name.
    >dim(data1)
    # [1] 475280 5
    I want to know what are the differentially expressed genes when treatment is used and if there is a species effect and a gene*species interaction and so on...

    >glmer.nbIA <- glmer.nb(counts~gene+treatment+species+gene*species+(1|animal)-1,data=data1, verbose=TRUE)

    But, the model does not run:
    >Error: cannot allocate vector of size 126.2 Gb

    I thought glmer.nb was adapted to deal with RNAseq data but maybe I am wrong. I did not find any publication using it on RNAseq data with lists of differentially expressed genes.
    Should the needed memory needed so huge?
    Thanks a lot for your help
    Gwenola Tosser-Klopp

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