In 20 µl reactions, recombinant She2p and She3p at indicated amounts were incubated with 5 nM 32 P-labeled ASH1 -E3-51 RNA for 20 min at 20°C in Dot-Blot buffer (pH 7.9).
UV cross-linking was performed for 15 min on ice, using a Spectrolinker XL-1500 (Spectroline) with an average intensity of 2,500 µW/cm 2.