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  • WGS and RNA-seq integrated in analysis of expression

    I need to find some publications about an analysis of the diversity of gene expression based on WGS and RNA-Seq.

    I would like to sequent the genome and the transtriptome (a sample of liver tissue and muscle tissue) in three individuals of pig. So I will have a 6 samples in RNA-Seq and 3 samples of WGS. I would like to explain differences in expression. Research will be based on WGS and RNA-Seq. Do you know articles about similar issues? Or any other proposition how to use a WGS and RNA-Seq together to do research about expression analysis?

  • #2
    I don't know any articles but it sounds to me like you're going to have to use these steps:
    1. use the WGS to assemble the genome
    2. align RNA-Seq to the assembled genome using an RNA-Seq aligner like STAR
    3. use Stringtie or Cufflinks to estimate transcript models from the aligned reads for each RNA-Seq sample
    4. merge the individual transcriptome assemblies with cuffmerge (if you used cufflinks) or Stringtie --merge (if you used stringtie)
    5. quantify each RNA-Seq sample's expression against the merged annotation (there are many options for this)
    6. Do differential expression testing in R using one of the many programs (DESeq2, edgeR, etc).

    Is there a reason why you cannot just align and quantify your RNA-Seq against the current Pig genome and gene annotations (susScr3)? Regardless, as a control I'd recommend doing that first so you can sanity check the de-novo pipeline.
    /* Shawn Driscoll, Gene Expression Laboratory, Pfaff
    Salk Institute for Biological Studies, La Jolla, CA, USA */

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