Pus7_rep2
PolyA RNA was treated with (N-cyclohexyl-N-?-(4-methylmorpholinium) (CMC) in bicine buffer (0.17 M CMC in 50 mM bicine, pH 8.3, 4 mM EDTA, 7 M urea) at 42C for 20 min. Excess CMC was removed by ethanol precipitation. To remove CMC groups attached to G and U, the CMC-treated RNA was subjected to alkali hydrolysis with Na2CO3 (50 mM, pH 10. 4) at 37C for 4 h. The reacted RNA was recovered by phenol: chloroform extraction, and ethanol precipitation.