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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.

Original publication

DOI

10.1038/s41592-024-02168-y

Type

Journal article

Journal

Nat methods

Publication Date

03/2024

Volume

21

Pages

401 - 405

Keywords

High-Throughput Nucleotide Sequencing, Sequence Analysis, RNA, Nucleotides, Oligonucleotides, Polymerase Chain Reaction