Literature DB >> 31124709

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

Clarisse Marotz1, Anukriti Sharma2, Greg Humphrey1, Neil Gottel2, Christopher Daum3, Jack A Gilbert2, Emiley Eloe-Fadrosh3, Rob Knight1,4,5.   

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.

Entities:  

Keywords:  16S rRNA gene amplicon sequencing; PCR; microbiome; replicate PCR reactions

Mesh：

Substances：

Year:  2019        PMID： 31124709      PMCID： PMC7030937          DOI： 10.2144/btn-2018-0192

Source DB:  PubMed          Journal:  Biotechniques        ISSN： 0736-6205            Impact factor:   1.993


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3.  Evaluating bias of illumina-based bacterial 16S rRNA gene profiles.

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