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Journal article

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

Abstract:
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.
Publication status:
Published
Peer review status:
Peer reviewed

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Publisher copy:
10.1038/s41592-024-02168-y

Authors


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Institution:
University of Oxford
Division:
MSD
Department:
NDORMS
Sub department:
Botnar Research Centre
Role:
Author
ORCID:
0000-0002-1274-5080
More by this author
Institution:
University of Oxford
Division:
MSD
Department:
NDORMS
Sub department:
Botnar Research Centre
Role:
Author
ORCID:
0000-0003-3632-0084
More by this author
Institution:
University of Oxford
Division:
MSD
Department:
NDORMS
Sub department:
Botnar Research Centre
Role:
Author
ORCID:
0000-0003-0137-9094
More by this author
Role:
Author
ORCID:
0009-0001-7932-9436
More by this author
Role:
Author
ORCID:
0000-0002-8706-4079


Publisher:
Springer Nature
Journal:
Nature Methods More from this journal
Volume:
21
Issue:
3
Pages:
401–405
Publication date:
2024-02-05
Acceptance date:
2024-01-04
DOI:
EISSN:
1548-7105
ISSN:
1548-7091


Language:
English
Keywords:
Pubs id:
1616655
Local pid:
pubs:1616655
Deposit date:
2024-02-13

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