Triplicate PCR reactions for 16S rRNA gene amplicon sequencing are unnecessary.
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ABSTRACT:
SUBMITTER: Marotz C
PROVIDER: S-EPMC7030937 | biostudies-literature | 2019 Jul
REPOSITORIES: biostudies-literature
Publications
Triplicate PCR reactions for 16S rRNA gene amplicon sequencing are unnecessary.
Marotz Clarisse C Sharma Anukriti A Humphrey Greg G Gottel Neil N Daum Christopher C Gilbert Jack A JA Eloe-Fadrosh Emiley E Knight Rob R
BioTechniques 20190524 1
Conventional wisdom holds that PCR amplification for sequencing should employ pooled replicate reactions to reduce bias due to jackpot effects and chimera formation. However, modern amplicon data analysis employs methods that may be less sensitive to such artifacts. Here we directly compare results from single versus triplicate reactions for 16S amplicon sequencing and find no significant impact of adopting a less labor-intensive single-reaction protocol. ...[more]
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