A practical method for barcoding and size-trimming PCR templates for amplicon sequencing
Abstract
Sample barcoding facilitates the analysis of tens or even hundreds of samples in a single next-generation sequencing (NGS) run, but more efficient methods are needed for high-throughput barcoding and size-trimming of long PCR products. Here we present a two-step PCR approach for barcoding followed by pool shearing, adapter ligation, and 5' end selection for trimming sets of DNA templates of any size. Our new trimming method offers clear benefits for phylogenetic studies, since targeting exactly the same region maximizes the alignment and enables the use of operational taxonomic unit (OTU)-based algorithms.
Main Authors
Format
Articles
Research article
Published
2016
Series
Subjects
Publication in research information system
Publisher
Informa Healthcare
The permanent address of the publication
https://urn.fi/URN:NBN:fi:jyu-201811014600Käytä tätä linkitykseen.
Review status
Peer reviewed
ISSN
0736-6205
DOI
https://doi.org/10.2144/000114380
Language
English
Published in
BioTechniques
Citation
- Mäki, A., Rissanen, A., & Tiirola, M. (2016). A practical method for barcoding and size-trimming PCR templates for amplicon sequencing. BioTechniques, 60(2), 88-90. https://doi.org/10.2144/000114380
License
Copyright© The Author(s) 2016.