Thread: 16S triplicates
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Old 08-12-2016, 12:14 PM   #2
thermophile
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Location: CT

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You should amplify your sample more than once because the seqs that get amplified in the first few rounds of amplification will dominate the amplicion pool. Since this is random, more common seqs should be among the first amplified but sometimes they aren't. The more reactions you initialize the more often you should see the "true" most abundant seqs being the most abundant at the end of PCR.

To minimize costs, you can reduce reaction volumes.

Google "16s triplicate pcr" for lots of papers to cite.

*"true" is a very lofty goal for 16S surveys but lets just go with it)
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