title: 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.
authors: Department of Biological and Environmental Science, University of Jyväskylä, Finland.
authors: Anita
authors: A
authors: Mäki
authors: Department of Biological and Environmental Science, University of Jyväskylä, Finland.
authors: Antti J
authors: AJ
authors: Rissanen
authors: Department of Biological and Environmental Science, University of Jyväskylä, Finland.
authors: Marja
authors: M
authors: Tiirola
mesh_terms: D000465:Algorithms; D004247:DNA / Q000235:genetics / Q000378:metabolism; D058893:DNA Barcoding, Taxonomic / Q000379:methods*; D015723:Gene Library; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*
chemical_list: D004247:DNA
keywords: PCR; ligation; next-generation sequencing; polyclonality; primer