Manganese Ions Individually Alter the Reverse Transcription Signature of Modified Ribonucleosides.

ABSTRACT: Reverse transcription of RNA templates containing modified ribonucleosides transfers modification-related information as misincorporations, arrest or nucleotide skipping events to the newly synthesized cDNA strand. The frequency and proportion of these events, merged from all sequenced cDNAs, yield a so-called RT signature, characteristic for the respective RNA modification and reverse transcriptase (RT). While known for DNA polymerases in so-called error-prone PCR, testing of four different RTs by replacing Mg2+ with Mn2+ in reaction buffer revealed the immense influence of manganese chloride on derived RT signatures, with arrest rates on m1A positions dropping from 82% down to 24%. Additionally, we observed a vast increase in nucleotide skipping events, with single positions rising from

SUBMITTER: Kristen M 

PROVIDER: S-EPMC7466121 | biostudies-literature | 2020 Aug

REPOSITORIES: biostudies-literature