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Triplicate PCR reactions for 16S rRNA gene amplicon sequencing are unnecessary

Identifiers: SRA: ERP113817
BioProject: PRJEB31281
UCSDMI: qiita_sid_11176
University of California San Diego Microbiome Initiative: qiita_sid_11176
Study Type: 
Other
Abstract: 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.
Description: 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.
Center Project: Triplicate_16S_PCR_Test
External Link: /PUBMED:31124709

Related SRA data

Experiments:
937 ( 937 samples )
Runs:
937 (47.8Gbp; 26.7Gb)