AUTHOR=Fresnedo-Ramírez Jonathan , Yang Shanshan , Sun Qi , Karn Avinash , Reisch Bruce I. , Cadle-Davidson Lance 

TITLE=Computational Analysis of AmpSeq Data for Targeted, High-Throughput Genotyping of Amplicons

JOURNAL=Frontiers in Plant Science

VOLUME=Volume 10 - 2019

YEAR=2019

URL=https://www.frontiersin.org/journals/plant-science/articles/10.3389/fpls.2019.00599

DOI=10.3389/fpls.2019.00599

ISSN=1664-462X

ABSTRACT=Amplicon sequencing (AmpSeq) is a practical, intuitive strategy with a semi-automated computational pipeline for analysis of highly multiplexed PCR-derived sequences.  This genotyping platform is particularly cost-effective when multiplexing 96 or more samples with a few amplicons up to thousands of amplicons. Amplicons can target from a single nucleotide to the upper limit of the sequencing platform. The flexibility of AmpSeq’s wet lab methods make it a tool of broad interest for diverse species, and AmpSeq excels in flexibility, high-throughput, low-cost, accuracy, and semi-automated analysis.  Here we provide an open science framework procedure to output data out of an AmpSeq project, with emphasis on the bioinformatics pipeline to generate SNPs, haplotypes and presence/absence variants in a set of diverse genotypes. Open-access tutorial datasets with actual data and a containerization open source software instance are provided to enable training in each of these genotyping applications. The pipelines presented here should be applicable to analysis of various target-enriched (eg, amplicon or sequence capture) Illumina sequence data.