Fig. 1: Scheme of MePMe-seq (metabolic propargylation for methylation sequencing). | Nature Communications

Fig. 1: Scheme of MePMe-seq (metabolic propargylation for methylation sequencing).

From: MePMe-seq: antibody-free simultaneous m6A and m5C mapping in mRNA by metabolic propargyl labeling and sequencing

Fig. 1

a Metabolic labeling of cells with PSH leads to methionine adenosyl transferase (MAT)-catalyzed formation of SAM-analogue and propargylation of methyltransferase (MTase) target sites. b After cell lysis, poly(A)+ RNA is isolated and fragmented. c Propargylated fragments react with biotin azide in a copper-catalyzed azide-alkyne cycloaddition (CuAAC) and are enriched on streptavidin-coated magnetic beads (SA mag. beads). d On-bead reverse transcription (RT) terminates at modified sites. e Libraries for next generation sequencing (NGS) are prepared. Modified sites are detected as coverage drops with distinct termination profiles.

Back to article page