The transcriptome-wide mapping of 2-methylthio-N6-isopentenyladenosine at single-base resolution by ReACT-seq

Through chemoselective methylthio group bioconjugation, we introduce a new approach (redox activated chemical tagging sequencing, ReACT-seq) to detect ms2i6A transcriptome-wide at single-base resolution. Comparative enrichment data analysis of ReACT-seq for ReACT strategy treated and no treated E. coli total RNA. For samples E.coli-RTase_SS, E.coli-RTase_HIV-RT, and E.coli-RTase_MMuLV, the protocols used were almost same, expect the choice of reverse transcriptase in library construction. To assess whether diverse reverse transcriptases have influence on the reverse transcription stop rate, Superscript III, HIV reverse transcriptase and MMuLV were used in sample E.coli-RTase_SS, E.coli-RTase_HIV-RT, and E.coli-RTase_MMuLV, respectively. ReACT label strategy were used in all three samples but not DBCO-s-s-Biotin enrichment method. Other protocols were the same for all samples.