Yan B, Tzertzinis G, Schildkraut I, Ettwiller L.

Genome Res. 2021 Nov 23. doi:10.1101/gr.275784.121

Comprehensive determination of transcription start sites derived from all RNA polymerases using ReCappable-seq.

5 µg of total RNAs was decapped with the the yeast scavenger decapping enzyme (yDcpS), and recapped with vaccinia capping enzyme (VCE) and a biotinylated guanosine. Therefore the protocol enriches for capped, triphosphorylated, diphosphorylated, but not monophosphorylated RNAs. A control library made on RNA dephosphorylated with CIP was used to infer if a TSS is driven by Pol II or Pol III. The methyl-triphosphate cap of RN7SK resists to the CIP treatement and causes it to be incorrectly classified Pol II.