Triplicate PCR Reactions for 16S rRNA Gene Amplicon Sequencing are Unnecessary
Copyright policy )Triplicate PCR Reactions for 16S rRNA Gene Amplicon Sequencing are Unnecessary
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.
- University of California, San Francisco United States
- University of California, San Diego United States
- Joint Genome Institute United States
- University of California, San Diego United States
- University of Chicago United States
DNA, Bacterial, 16S, Technology, Bioinformatics, QH301-705.5, replicate PCR reactions, 610, microbiome, Polymerase Chain Reaction, 630, RNA, Ribosomal, 16S, Genetics, Biology (General), 16S rRNA gene amplicon sequencing, Ribosomal, Bacteria, Bacterial, DNA, Sequence Analysis, DNA, Biological Sciences, PCR, RNA, Sequence Analysis, Biotechnology
DNA, Bacterial, 16S, Technology, Bioinformatics, QH301-705.5, replicate PCR reactions, 610, microbiome, Polymerase Chain Reaction, 630, RNA, Ribosomal, 16S, Genetics, Biology (General), 16S rRNA gene amplicon sequencing, Ribosomal, Bacteria, Bacterial, DNA, Sequence Analysis, DNA, Biological Sciences, PCR, RNA, Sequence Analysis, Biotechnology
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