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

Helen M. Gunter, Scott E. Youlten, Bindu Swapna Madala, Andre L. M. Reis, Igor Stevanovski, Ted Wong, Sarah K. Kummerfield, Ira W. Deveson, Nadia S. Santini, Esteban Marcellin and Tim R. Mercer ()
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Helen M. Gunter: University of Queensland
Scott E. Youlten: Garvan Institute of Medical Research
Bindu Swapna Madala: Garvan Institute of Medical Research
Andre L. M. Reis: Garvan Institute of Medical Research
Igor Stevanovski: Garvan Institute of Medical Research
Ted Wong: Garvan Institute of Medical Research
Sarah K. Kummerfield: Garvan Institute of Medical Research
Ira W. Deveson: Garvan Institute of Medical Research
Nadia S. Santini: Instituto de Ecología, Universidad Nacional Autónoma de México
Esteban Marcellin: University of Queensland
Tim R. Mercer: University of Queensland

Nature Communications, 2022, vol. 13, issue 1, 1-11

Abstract: Abstract 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.

Date: 2022
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DOI: 10.1038/s41467-022-34028-8

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