Normalizing large scale RNA-seq datasets
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zqzneptune • 0
@zqzneptune-12327
Last seen 7.1 years ago

I am currently processing large scale RNA-seq data from independent cohorts.

Is there a systematic strategy to normalize the TPM of each transcript so that the expression levels can be compared without concerning the batch effect?

Thanks!

 

deseq2 • 1.2k views
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@mikelove
Last seen 5 hours ago
United States

A critical question here is if you are interested in making comparisons within cohorts (e.g. comparing disease vs normal across numerous studies, each with normal and disease samples), or across cohorts (e.g. this study has disease A, this other study has disease B). The first kind of comparison is called "balanced" while the second one is "confounded".

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The problem here tends to be the first scenario, where samples from the tissues are to be compared to generate hypothesis. Since, this is a "balanced" case, is the package "sva" one of the solutions?

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Yes, sva is appropriate and a great normalization tool. You can make contrasts in limma as well after sva normalization. 

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