I have seen some papers on pooling and apparently it's a statical mouth watering conversation so let me explain how this "pooling" would be done before anyone raises hell.
The experiment is RNA-seq to determine differentially expressed genes between wild type bacterial strain and a mutant. Call them WT and M
You need three biological replicates for both WT and M. Let's talk about just WT for now because it is the same for M.
What you do is take three colonies from WT and grow overnight cultures. Call them WT1, WT2, WT3.
Next day take the three overnight culture and reinnoculate them but this time make two culture for each overnight culture so you end up with six total where only three are from different colonies.
Call them WT1(a), WT1(b), WT2(a), WT2(b), WT3(a), WT3(b)
Grow them to a desired OD and then pool WT1(a+b), WT2(a+b), WT3(a+b) so you end up with three biological replicates coming from two independent growths.
What are your thoughts on making biological replicates this way?
The experiment is RNA-seq to determine differentially expressed genes between wild type bacterial strain and a mutant. Call them WT and M
You need three biological replicates for both WT and M. Let's talk about just WT for now because it is the same for M.
What you do is take three colonies from WT and grow overnight cultures. Call them WT1, WT2, WT3.
Next day take the three overnight culture and reinnoculate them but this time make two culture for each overnight culture so you end up with six total where only three are from different colonies.
Call them WT1(a), WT1(b), WT2(a), WT2(b), WT3(a), WT3(b)
Grow them to a desired OD and then pool WT1(a+b), WT2(a+b), WT3(a+b) so you end up with three biological replicates coming from two independent growths.
What are your thoughts on making biological replicates this way?
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