Systematic evaluation of parameters in RNA bisulfite sequencing data generation and analysis.

ABSTRACT: The presence of 5-methylcytosine (m⁵C) in RNA molecules has been known for decades and its importance in regulating RNA metabolism has gradually become appreciated. Despite recent advances made in the functional and mechanistic understanding of RNA m⁵C modifications, the detection and quantification of methylated RNA remains a challenge. In this study, we compared four library construction procedures for RNA bisulfite sequencing and implemented an analytical pipeline to assess the key parameters in the process of m⁵C calling. We found that RNA fragmentation after bisulfite conversion increased the yield significantly, and an additional high temperature treatment improved bisulfite conversion efficiency especially for sequence reads mapped to the mitochondri

SUBMITTER: Johnson Z 

PROVIDER: S-EPMC9164272 | biostudies-literature | 2022 Jun

REPOSITORIES: biostudies-literature