I often see in studies that whenever people prepare samples for 16S sequencing there's often a line which says that each sample was amplified in triplicates.
Why?
Is there any paper out there which explains the need for having 16S samples in triplicates? I couldn't find anything informative...
I understand why somebody would make triplicates in cases where the amplification for that sample would fail due to technical issues. You'd end up with less samples and have to repeat some maybe, but from an analysis point of view, is there really a need?
Is it to potentially average out the variation caused during amplification or sequencing? Any papers showing this effect?
It's not an issue if your sample size is small, but if you are dealing with 1000s of samples, it does get quite costly in terms of PCR/adapter oligo synthesis and PCR mix quantities.
Just trying to avoid unnecessary cost...
Why?
Is there any paper out there which explains the need for having 16S samples in triplicates? I couldn't find anything informative...
I understand why somebody would make triplicates in cases where the amplification for that sample would fail due to technical issues. You'd end up with less samples and have to repeat some maybe, but from an analysis point of view, is there really a need?
Is it to potentially average out the variation caused during amplification or sequencing? Any papers showing this effect?
It's not an issue if your sample size is small, but if you are dealing with 1000s of samples, it does get quite costly in terms of PCR/adapter oligo synthesis and PCR mix quantities.
Just trying to avoid unnecessary cost...
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