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. 1997 Sep 30;94(20):10612–10617. doi: 10.1073/pnas.94.20.10612

Figure 2.

Figure 2

(A) Self-cleavage of clone 8 RNA in the different buffer systems studied. Internally labeled RNA was incubated at room temperature in TE buffer at pH 7.5 (lane 1), 50 mM sodium acetate buffer at pH 4.8 (lane 2), 50 mM ammonium acetate buffer at pH 4.8 (lane 3), 50 mM sodium citrate at pH 5.0 (lane 4), 50 mM sodium Mes buffer at pH 6.7 (lane 5) or pH 5.5 (lane 6), sodium cacodylate buffer at pH 7.4 (lane 7) or pH 5.0 (lane 8), potassium phosphate buffer at pH 8.0 (lane 9) or pH 5.8 (lane 10) at a final RNA concentration of 1 μM (15). (B) Site of self-cleavage for clone 8 RNA. RNA labeled at the 5′ end was treated with 50 mM sodium cacodylate buffer at pH 7.4 (lane 1) or pH 5.0 (lane 2). To determine the cleavage site, samples were run on a 10% polyacrylamide/7 M urea gel along with partial alkaline hydrolysis ladder (lane 3) and partial RNase T1 digestion samples (lane 4). (C) Effect of pH on cleavage of clone 8 RNA in 50 mM sodium citrate buffer. The rate of cleavage obtained at each pH value was normalized to the maximum rate measured at pH 4.2.