Elimination of PCR duplicates in RNA-seq and small RNA-seq using unique molecular identifiers.

ABSTRACT:

Background

RNA-seq and small RNA-seq are powerful, quantitative tools to study gene regulation and function. Common high-throughput sequencing methods rely on polymerase chain reaction (PCR) to expand the starting material, but not every molecule amplifies equally, causing some to be overrepresented. Unique molecular identifiers (UMIs) can be used to distinguish undesirable PCR duplicates derived from a single molecule and identical but biologically meaningful reads from different molecules.

Results

We have incorporated UMIs into RNA-seq and small RNA-seq protocols and developed tools to analyze the resulting data. Our UMIs contain stretches of random nucleotides whose lengths sufficiently capture diverse molecule species in both RNA-seq and small RNA-seq libraries generated

SUBMITTER: Fu Y 

PROVIDER: S-EPMC6044086 | biostudies-literature | 2018 Jul

REPOSITORIES: biostudies-literature