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Journal of Biomolecular Techniques : JBT logoLink to Journal of Biomolecular Techniques : JBT
. 2019 Dec;30(Suppl):S35–S36.

RipTide High Throughput NGS Library Prep for Genotyping in Populations

Azeem Siddique 1,, Gaia Suckow 2, Phillip Ordoukhanian 2, Steve Head 2, Nils Homer 3, Alvaro Hernandez 4, Keith Brown 1, Lior Glick 5, Kobi Baruch 5, Paul Doran 1
PMCID: PMC6936909

Abstract

High throughput genotyping technologies are required for large-scale population genetics. Evolutionary biology studies, human disease research and large-scale agricultural breeding programs all lend themselves to technologies that are able to provide more information at lower cost. Over the past decade, genotyping technology has transitioned from PCR-based SNP assays to microarrays, and is now shifting toward high-throughput genotyping by sequencing (GBS). The RipTide High Throughput Rapid DNA Library Prep allows for the preparation of NGS libraries from up to 960 individually barcoded samples in a few hours with automation. When combined with low coverage sequencing and imputation-based genotype analysis, the result is an order of magnitude greater information at a significantly reduced cost. Here we present data on 96 Zea mays (maize) samples consisting of 4 parent populations and 92 recombinant inbred lines (RILs). For each sample, hundreds of thousands to millions of haplotype markers, including SNVs and structural variants, are accurately detected. A minimum of 95% complete coverage of direct and imputed markers is obtained for each RIL. The approach can be applied to any species, regardless of genome size or GC content. In this study, a median of >1 million markers were genotyped by sequencing on an Illumina HiSeq 4000 instrument for an estimated cost of library construction and sequencing of < $25 per sample.


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