How may libraries should I consider?
Hi everyone I need help. I start working on RNAseq and I'd like to know how many libraries should I consider for my experiment. I'm working on the fish gut microbiome and I'm trying to extract the RNA from the feces. I have 60 samples to analyse.
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Each sample will be one library. Or did I misunderstand your question?
I mean is one library enough for each sample? Or should I consider more than one?
Yes, one library is sufficient. RNA-seq has generally good enough technical replicability, so you should focus on biological replicates (include more individuals in your study for proper statistics) and not technical replicates (more libraries from the same individual).
Ok...thank you for your help!
What do you hope to get from the analysis? If this is meta-transcriptomic data you would obviously not be doing conventional differential expression analysis.
It`s an exposure experiment to different oil conc...we have already seen significant difference in the microbial community but now we want to understand what does it really mean...