6533b835fe1ef96bd129f466
RESEARCH PRODUCT
A practical method for barcoding and size-trimming PCR templates for amplicon sequencing
Anita MäkiAntti J. RissanenMarja Tiirolasubject
0301 basic medicineOperational taxonomic unitComputer science030106 microbiologyLong pcrComputational biologyPolymerase Chain ReactionGeneral Biochemistry Genetics and Molecular BiologyDNA sequencinglaw.invention03 medical and health scienceslawDNA Barcoding TaxonomicGenomic libraryligationPolymerase chain reactionGene Libraryta1184ta1182High-Throughput Nucleotide SequencingDNAMolecular biologyprimer030104 developmental biologyTemplatePCRpolyclonalityAmplicon sequencingTrimmingnext-generation sequencingAlgorithmsBiotechnologydescription
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.
| year | journal | country | edition | language |
|---|---|---|---|---|
| 2016-01-01 |