Triplicate PCR reactions for 16S rRNA gene amplicon sequencing are unnecessary.
16S rRNA gene amplicon sequencing
PCR
microbiome
replicate PCR reactions
Journal
BioTechniques
ISSN: 1940-9818
Titre abrégé: Biotechniques
Pays: England
ID NLM: 8306785
Informations de publication
Date de publication:
07 2019
07 2019
Historique:
pubmed:
28
5
2019
medline:
19
5
2020
entrez:
25
5
2019
Statut:
ppublish
Résumé
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.
Identifiants
pubmed: 31124709
doi: 10.2144/btn-2018-0192
pmc: PMC7030937
mid: NIHMS1064766
doi:
Substances chimiques
DNA, Bacterial
0
RNA, Ribosomal, 16S
0
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, U.S. Gov't, Non-P.H.S.
Langues
eng
Sous-ensembles de citation
IM
Pagination
29-32Subventions
Organisme : NIDCR NIH HHS
ID : F31 DE028478
Pays : United States
Références
Nat Biotechnol. 2017 Nov;35(11):1077-1086
pubmed: 28967885
Appl Environ Microbiol. 2014 Sep;80(18):5717-22
pubmed: 25002428
Appl Environ Microbiol. 1998 Oct;64(10):3724-30
pubmed: 9758791
Biotechniques. 2017 Jun 1;62(6):290-293
pubmed: 28625159
Genome Biol. 2013 Feb 28;14(2):202
pubmed: 23514020
PLoS One. 2017 Apr 28;12(4):e0176716
pubmed: 28453559
J Vis Exp. 2018 Jul 24;(137):
pubmed: 30102263
Nat Biotechnol. 2019 Aug;37(8):852-857
pubmed: 31341288
mSystems. 2017 Mar 7;2(2):
pubmed: 28289731
mSystems. 2018 Nov 6;3(6):
pubmed: 30417111
mSystems. 2018 May 29;3(3):
pubmed: 29854953
PLoS One. 2014 Feb 28;9(2):e90234
pubmed: 24587293