A neutral pH thermal hydrolysis method for quantification of structured RNAs

Neutral thermal hydrolysis or quantitative NMR can be used to determine RNA concentrations. (A) Time course of the neutral thermal hydrolysis reactions for different riboswitch aptamers. In each case, the 120-min time point was taken as the A_{260,hydrolyzed} value to calculate the RNA concentration shown in Table 1. (B) ³¹P-NMR spectrum for a nonhydrolyzed RNA sample (the SAM-I riboswitch aptamer upstream of the PI23P_02107 gene from Polaribacter irgensii, Pi). The same RNA sample was analyzed by neutral thermal hydrolysis and the results are shown in Table 1.