Library adaptors with integrated reference controls improve the accuracy and reliability of nanopore sequencing.

Autor: Gunter HM; Australian Institute of Bioengineering and Nanotechnology, University of Queensland, Brisbane, QLD, Australia., Youlten SE; Kinghorn Centre for Clinical Genomics, Garvan Institute of Medical Research, Sydney, NSW, Australia., Madala BS; Kinghorn Centre for Clinical Genomics, Garvan Institute of Medical Research, Sydney, NSW, Australia., Reis ALM; Kinghorn Centre for Clinical Genomics, Garvan Institute of Medical Research, Sydney, NSW, Australia., Stevanovski I; Kinghorn Centre for Clinical Genomics, Garvan Institute of Medical Research, Sydney, NSW, Australia., Wong T; Kinghorn Centre for Clinical Genomics, Garvan Institute of Medical Research, Sydney, NSW, Australia., Kummerfield SK; Kinghorn Centre for Clinical Genomics, Garvan Institute of Medical Research, Sydney, NSW, Australia.; St Vincent's Clinical School, Faculty of Medicine, University of New South Wales, Sydney, NSW, Australia., Deveson IW; Kinghorn Centre for Clinical Genomics, Garvan Institute of Medical Research, Sydney, NSW, Australia.; St Vincent's Clinical School, Faculty of Medicine, University of New South Wales, Sydney, NSW, Australia., Santini NS; Programa Investigadoras e Investigadores por México, Instituto de Ecología, Universidad Nacional Autónoma de México, Mexico City, Mexico., Marcellin E; Australian Institute of Bioengineering and Nanotechnology, University of Queensland, Brisbane, QLD, Australia., Mercer TR; Australian Institute of Bioengineering and Nanotechnology, University of Queensland, Brisbane, QLD, Australia. t.mercer@uq.edu.au.; Kinghorn Centre for Clinical Genomics, Garvan Institute of Medical Research, Sydney, NSW, Australia. t.mercer@uq.edu.au.
Jazyk: angličtina
Zdroj: Nature communications [Nat Commun] 2022 Oct 28; Vol. 13 (1), pp. 6437. Date of Electronic Publication: 2022 Oct 28.
DOI: 10.1038/s41467-022-34028-8
Abstrakt: Library adaptors are short oligonucleotides that are attached to RNA and DNA samples in preparation for next-generation sequencing (NGS). Adaptors can also include additional functional elements, such as sample indexes and unique molecular identifiers, to improve library analysis. Here, we describe Control Library Adaptors, termed CAPTORs, that measure the accuracy and reliability of NGS. CAPTORs can be integrated within the library preparation of RNA and DNA samples, and their encoded information is retrieved during sequencing. We show how CAPTORs can measure the accuracy of nanopore sequencing, evaluate the quantitative performance of metagenomic and RNA sequencing, and improve normalisation between samples. CAPTORs can also be customised for clinical diagnoses, correcting systematic sequencing errors and improving the diagnosis of pathogenic BRCA1/2 variants in breast cancer. CAPTORs are a simple and effective method to increase the accuracy and reliability of NGS, enabling comparisons between samples, reagents and laboratories, and supporting the use of nanopore sequencing for clinical diagnosis.
(© 2022. The Author(s).)
Databáze: MEDLINE
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