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ERX3190095: Illumina MiSeq sequencing; qiita_ptid_6188:11176.1x.CC0114F.E7.072516.186
1 ILLUMINA (Illumina MiSeq) run: 464,657 spots, 123.4M bases, 79.7Mb downloads

Design: comparison of triplicate vs single PCR reactions
Submitted by: University of California San Diego Microbiome Initiative (University of California San Diego Microbiome Init)
Study: Triplicate PCR reactions for 16S rRNA gene amplicon sequencing are unnecessary
show Abstracthide 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.
Sample: triplicate_test; 11176.1x.CC0114F.E7.072516.186
SAMEA5358117 • ERS3163738 • All experiments • All runs
Library:
Name: 11176.1x.CC0114F.E7.072516.186
Instrument: Illumina MiSeq
Strategy: OTHER
Source: METAGENOMIC
Selection: PCR
Layout: SINGLE
Construction protocol: Illumina EMP protocol 515fbc, 806r amplification of 16S rRNA V4
Experiment attributes: (show all 14 attributes...) (hide...)
barcode: ATGCGAGTATC
center_name: JGI
center_project_name: JGI triplicate testing
direction: forward
forward_read: 11514.1.209094.ATGCGAGTATC.fastq.R1.fastq.gz
linker: GT
pcr_primers: FWD:GTGYCAGCMGCCGCGGTAA; REV:GGACTACNVGGGTWTCTAAT
primer: GTGTGYCAGCMGCCGCGGTAA
run_center: JGI
run_date: 2017
run_prefix: 11514.1.209094.ATGCGAGTATC
sequencing_meth: Sequencing by synthesis
target_gene: 16S rRNA
target_subfragment: V4
Runs: 1 run, 464,657 spots, 123.4M bases, 79.7Mb
Run# of Spots# of BasesSizePublished
ERR3162011464,657123.4M79.7Mb2020-02-07

ID:
10041658

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