Preparation of p53 Variant RNA Templates

The dsDNA templates for the preparation of P0-Δ40p53, P1-Δ40p53, P1-Δ40p53(L), P1-Δ40p53(S), P1-Δ40p53(Δ57), P1-Δ40p53(ΔHDM2), p53-554 and isolated hairpin G56-C169 RNAs had been obtained previously in our laboratory [38 (link),39 (link),63 (link)]. For transcription in vitro, the dsDNA templates were first linearised with Csp45 or Xba1 restriction enzymes (Thermo Fisher, Walthan, MA, USA) in accordance with the manufacturer’s protocols. Transcription reactions were performed using the TranscriptAid T7 High Yield Transcription Kit (Thermo Fisher, Walthan, MA, USA) as recommended by the manufacturer’s protocol and with an addition of 4 mM guanosine. After the transcription, RNAs were purified using the GeneJet RNA Isolation Kit (Thermo Fisher, Walthan, MA, USA).