Correcting PCR amplification errors in unique molecular identifiers to generate accurate numbers of sequencing molecules
Jianfeng Sun, Martin Philpott, Danson Loi, Shuang Li, Pablo Monteagudo-Mesas, Gabriela Hoffman, Jonathan Robson, Neelam Mehta, Vicki Gamble, Tom Brown, Tom Brown, Stefan Canzar, Udo Oppermann, Adam P. Cribbs
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.
Topics & Concepts
IdentifierOligonucleotideComputational biologyBiologyComputer scienceGeneticsDNAProgramming languageMolecular Biology Techniques and ApplicationsEnvironmental DNA in Biodiversity StudiesCancer Genomics and Diagnostics