NanoGBS: A Miniaturized Procedure for GBS Library Preparation.
GBS library construction
NanoGBS
genotyping
genotyping-by-sequencing
reduced-representation sequencing
Journal
Frontiers in genetics
ISSN: 1664-8021
Titre abrégé: Front Genet
Pays: Switzerland
ID NLM: 101560621
Informations de publication
Date de publication:
2020
2020
Historique:
received:
04
10
2019
accepted:
20
01
2020
entrez:
6
3
2020
pubmed:
7
3
2020
medline:
7
3
2020
Statut:
epublish
Résumé
High-throughput reduced-representation sequencing (RRS)-based genotyping methods, such as genotyping-by-sequencing (GBS), have provided attractive genotyping solutions in numerous species. Here, we present NanoGBS, a miniaturized and eco-friendly method for GBS library construction. Using acoustic droplet ejection (ADE) technology, NanoGBS libraries were constructed in tenfold smaller volumes compared to standard methods (StdGBS) and leading to a reduced use of plastics of up to 90%. A high-quality DNA library and SNP catalogue were obtained with extensive overlap (96%) in SNP loci and 100% agreement in genotype calls compared to the StdGBS dataset with a high level of accuracy (98.5%). A highly multiplexed pool of GBS libraries (768-plex) was sequenced on a single Ion Proton PI chip and yielded enough SNPs (~4K SNPs; 1.5 SNP per cM, on average) for many high-volume applications. Combining NanoGBS library preparation and increased multiplexing can dramatically reduce (72%) genotyping cost per sample. We believe that this approach will greatly facilitate the adoption of marker applications where extremely high throughputs are required and cost is still currently limiting.
Identifiants
pubmed: 32133028
doi: 10.3389/fgene.2020.00067
pmc: PMC7040475
doi:
Types de publication
Journal Article
Langues
eng
Pagination
67Informations de copyright
Copyright © 2020 Torkamaneh, Boyle, St-Cyr, Légaré, Pomerleau and Belzile.
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