Library adaptors with integrated reference controls improve the accuracy and reliability of nanopore sequencing
Por:
Gunter, Helen M., Youlten, Scott E., Madala, Bindu Swapna, Reis A.L.M., Stevanovski, Igor, Wong, Ted, Kummerfield, Sarah K., Deveson, Ira W., Santini, Nadia S., Marcellin, Esteban, Mercer, Tim R.
Publicada:
1 ene 2022
Resumen:
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).
Filiaciones:
Gunter, Helen M.:
Australian Institute of Bioengineering and Nanotechnology, University of Queensland, Brisbane, QLD, Australia
Youlten, Scott E.:
Kinghorn Centre for Clinical Genomics, Garvan Institute of Medical Research, Sydney, NSW, Australia
Madala, Bindu Swapna:
Kinghorn Centre for Clinical Genomics, Garvan Institute of Medical Research, Sydney, NSW, Australia
Reis A.L.M.:
Kinghorn Centre for Clinical Genomics, Garvan Institute of Medical Research, Sydney, NSW, Australia
Stevanovski, Igor:
Kinghorn Centre for Clinical Genomics, Garvan Institute of Medical Research, Sydney, NSW, Australia
Wong, Ted:
Kinghorn Centre for Clinical Genomics, Garvan Institute of Medical Research, Sydney, NSW, Australia
Kummerfield, Sarah K.:
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, Ira W.:
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, Nadia S.:
Programa Investigadoras e Investigadores por México, Instituto de Ecología, Universidad Nacional Autónoma de México, Mexico City, Mexico
Marcellin, Esteban:
Australian Institute of Bioengineering and Nanotechnology, University of Queensland, Brisbane, QLD, Australia
Mercer, Tim R.:
Australian Institute of Bioengineering and Nanotechnology, University of Queensland, Brisbane, QLD, Australia
Kinghorn Centre for Clinical Genomics, Garvan Institute of Medical Research, Sydney, NSW, Australia
gold, Green Published, All Open Access, Gold, Green
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