Question: How may libraries should I consider?
0
gravatar for Giovy
4 weeks ago by
Giovy0
Giovy0 wrote:

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.

library rna-seq • 100 views
ADD COMMENTlink modified 4 weeks ago by h.mon28k • written 4 weeks ago by Giovy0

Each sample will be one library. Or did I misunderstand your question?

ADD REPLYlink written 4 weeks ago by WouterDeCoster42k

I mean is one library enough for each sample? Or should I consider more than one?

ADD REPLYlink written 4 weeks ago by Giovy0

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).

ADD REPLYlink written 4 weeks ago by WouterDeCoster42k

Ok...thank you for your help!

ADD REPLYlink written 4 weeks ago by Giovy0

I'm working on the fish gut microbiome and I'm trying to extract the RNA from the feces.

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.

ADD REPLYlink modified 4 weeks ago • written 4 weeks ago by genomax75k

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...

ADD REPLYlink modified 4 weeks ago • written 4 weeks ago by Giovy0
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