Insufficiently complex unique-molecular identifiers (UMIs) distort small RNA sequencing.

ABSTRACT: The attachment of unique molecular identifiers (UMIs) to RNA molecules prior to PCR amplification and sequencing, makes it possible to amplify libraries to a level that is sufficient to identify rare molecules, whilst simultaneously eliminating PCR bias through the identification of duplicated reads. Accurate de-duplication is dependent upon a sufficiently complex pool of UMIs to allow unique labelling. In applications dealing with complex libraries, such as total RNA-seq, only a limited variety of UMIs are required as the variation in molecules to be sequenced is enormous. However, when sequencing a less complex library, such as small RNAs for which there is a more limited range of possible sequences, we find increased variation in UMIs are required, even beyond that provided in a commerc

SUBMITTER: Saunders K 

PROVIDER: S-EPMC7471316 | biostudies-literature | 2020 Sep

REPOSITORIES: biostudies-literature