Correcting PCR amplification errors in unique molecular identifiers to generate accurate numbers of sequencing molecules.

Autor: Sun J; Botnar Research Centre, Nuffield Department of Orthopaedics, Rheumatology and Musculoskeletal Sciences, National Institute of Health Research Oxford Biomedical Research Unit (BRU), University of Oxford, Oxford, UK., Philpott M; Botnar Research Centre, Nuffield Department of Orthopaedics, Rheumatology and Musculoskeletal Sciences, National Institute of Health Research Oxford Biomedical Research Unit (BRU), University of Oxford, Oxford, UK., Loi D; Botnar Research Centre, Nuffield Department of Orthopaedics, Rheumatology and Musculoskeletal Sciences, National Institute of Health Research Oxford Biomedical Research Unit (BRU), University of Oxford, Oxford, UK., Li S; Gene Center, Ludwig-Maximilians University of Munich, Munich, Germany., Monteagudo-Mesas P; Gene Center, Ludwig-Maximilians University of Munich, Munich, Germany., Hoffman G; ATDBio Ltd (now part of Biotage), Magdalen Centre, Oxford Science Park, Oxford, UK., Robson J; ATDBio Ltd (now part of Biotage), Magdalen Centre, Oxford Science Park, Oxford, UK., Mehta N; Botnar Research Centre, Nuffield Department of Orthopaedics, Rheumatology and Musculoskeletal Sciences, National Institute of Health Research Oxford Biomedical Research Unit (BRU), University of Oxford, Oxford, UK., Gamble V; Botnar Research Centre, Nuffield Department of Orthopaedics, Rheumatology and Musculoskeletal Sciences, National Institute of Health Research Oxford Biomedical Research Unit (BRU), University of Oxford, Oxford, UK., Brown T Jr; ATDBio Ltd (now part of Biotage), Magdalen Centre, Oxford Science Park, Oxford, UK., Brown T Sr; Chemistry Research Laboratory, Department of Chemistry, University of Oxford, Oxford, UK., Canzar S; Department of Computer Science and Engineering, The Pennsylvania State University, University Park, PA, USA.; Huck Institutes of the Life Sciences, The Pennsylvania State University, University Park, PA, USA., Oppermann U; Botnar Research Centre, Nuffield Department of Orthopaedics, Rheumatology and Musculoskeletal Sciences, National Institute of Health Research Oxford Biomedical Research Unit (BRU), University of Oxford, Oxford, UK.; Oxford Centre for Translational Myeloma Research, University of Oxford, Oxford, UK., Cribbs AP; Botnar Research Centre, Nuffield Department of Orthopaedics, Rheumatology and Musculoskeletal Sciences, National Institute of Health Research Oxford Biomedical Research Unit (BRU), University of Oxford, Oxford, UK. adam.cribbs@ndorms.ox.ac.uk.; Oxford Centre for Translational Myeloma Research, University of Oxford, Oxford, UK. adam.cribbs@ndorms.ox.ac.uk.
Jazyk: angličtina
Zdroj: Nature methods [Nat Methods] 2024 Mar; Vol. 21 (3), pp. 401-405. Date of Electronic Publication: 2024 Feb 05.
DOI: 10.1038/s41592-024-02168-y
Abstrakt: Unique molecular identifiers are random oligonucleotide sequences that remove PCR amplification biases. However, the impact that PCR associated sequencing errors have on the accuracy of generating absolute counts of RNA molecules is underappreciated. We show that PCR errors are a source of inaccuracy in both bulk and single-cell sequencing data, and synthesizing unique molecular identifiers using homotrimeric nucleotide blocks provides an error-correcting solution that allows absolute counting of sequenced molecules.
(© 2024. The Author(s).)
Databáze: MEDLINE
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