Accurate characterization of Escherichia coli tRNA modifications with a simple method of deep-sequencing library preparation.

ABSTRACT: In conventional RNA high-throughput sequencing, modified bases prevent a large fraction of tRNA transcripts to be converted into cDNA libraries. Recent proposals aiming at resolving this issue take advantage of the interference of base modifications with RT enzymes to detect and identify them by establishing signals from aborted cDNA transcripts. Because some modifications, such as methyl groups, do almost not allow RT bypassing, demethylation and highly processive RT enzymes have been used to overcome these obstacles. Working with Escherichia coli as a model system, we show that with a conventional (albeit still engineered) RT enzyme and key optimizations in library preparation, all RT-impairing modifications can be highlighted along the entire tRNA length without demethylation pro

SUBMITTER: Wang J 

PROVIDER: S-EPMC7833731 | biostudies-literature | 2021 Jan

REPOSITORIES: biostudies-literature