Practical innovations for high-throughput amplicon sequencing

A set of practical improvements and software provide more accurate and less biased metagenomic amplicon sequencing at lower sequencing effort. We describe improvements for sequencing 16S ribosomal RNA (rRNA) amplicons, a cornerstone technique in metagenomics. Through unique tagging of template molec...

Full description

Saved in:
Bibliographic Details
Published inNature methods Vol. 10; no. 10; pp. 999 - 1002
Main Authors Lundberg, Derek S, Yourstone, Scott, Mieczkowski, Piotr, Jones, Corbin D, Dangl, Jeffery L
Format Journal Article
LanguageEnglish
Published New York Nature Publishing Group US 01.10.2013
Nature Publishing Group
Subjects
631/1647/514/1948
631/208/212/2142
631/326/2565/2142
Bacteria - classification
Bacteria - genetics
Bioinformatics
Biological Microscopy
Biological Techniques
Biomedical Engineering/Biotechnology
brief-communication
Classification - methods
Deoxyribonucleic acid
DNA
DNA sequencing
Genomics
High-Throughput Nucleotide Sequencing - methods
Life Sciences
Metagenome
Methods
Nucleotide sequencing
Polymerase chain reaction
Proteomics
RNA, Ribosomal, 16S - genetics
Software
Soil Microbiology
United States
Online AccessGet full text

Cover

More Information
Summary:A set of practical improvements and software provide more accurate and less biased metagenomic amplicon sequencing at lower sequencing effort. We describe improvements for sequencing 16S ribosomal RNA (rRNA) amplicons, a cornerstone technique in metagenomics. Through unique tagging of template molecules before PCR, amplicon sequences can be mapped to their original templates to correct amplification bias and sequencing error with software we provide. PCR clamps block amplification of contaminating sequences from a eukaryotic host, thereby substantially enriching microbial sequences without introducing bias.
Bibliography:ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
content type line 23
ISSN:1548-7091
1548-7105
DOI:10.1038/nmeth.2634