Unique Molecular Identifiers reveal a novel sequencing artefact with implications for RNA-Seq based gene expression analysis.

ABSTRACT: Attaching Unique Molecular Identifiers (UMI) to RNA molecules in the first step of sequencing library preparation establishes a distinct identity for each input molecule. This makes it possible to eliminate the effects of PCR amplification bias, which is particularly important where many PCR cycles are required, for example, in single cell studies. After PCR, molecules sharing a UMI are assumed to be derived from the same input molecule. In our single cell RNA-Seq studies of Physcomitrella patens, we discovered that reads sharing a UMI, and therefore presumed to be derived from the same mRNA molecule, frequently map to different, but closely spaced locations. This behaviour occurs in all such libraries that we have produced, and in multiple other UMI-containing RNA-Seq data sets in the pub

SUBMITTER: Sena JA 

PROVIDER: S-EPMC6120941 | biostudies-literature | 2018 Sep

REPOSITORIES: biostudies-literature