Hello all,
I plan to work on microbiome. We have 20 controls and 20 patient. Our budget is limited and so we intend to pool the samples. My question is that whether we can pool samples before library prep or pooling samples is just after library-prep? By the way, does anyone know pooling might be statistically problematic? how about the library-prep kit? can you name some of them to be appropriate for our sample size? Any thought will be appreciated
1 answer
You have to prep the libraries separately with appropriate indexes (if you want to be able to distinguish reads coming from specific samples). You will then pool the prepared libraries before sequencing them as a pool. You have to quantify/QC the libraries independently before deciding on a pooling scheme. The pool is generally quantified and QC'ed as well.
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