An integrative protocol for one-step PCR amplicon library construction and accurate demultiplexing of pooled sequencing data.

ABSTRACT: High-throughput sequencing of amplicons has been widely used to precisely and efficiently identify species compositions and analyze community structures, greatly promoting biological studies involving large amounts of complex samples, especially those involving environmental and pathogen-monitoring ones. Commercial library preparation kits for amplicon sequencing, which generally require multiple steps, including adapter ligation and indexing, are expensive and time-consuming, especially for applications at a large scale. To overcome these limitations, a "one-step PCR approach" has been previously proposed for constructions of amplicon libraries using long fusion primers. However, efficient amplifications of target genes and accurate demultiplexing of pooled sequencing data remain to be ad

SUBMITTER: Ni J 

PROVIDER: S-EPMC10689312 | biostudies-literature | 2023 Nov

REPOSITORIES: biostudies-literature