Figure 6.
Effect of the primer length on the polymerization-dependent RNase H cleavage patterns obtained with WT BH10 and 3M_T355A/Q357M RTs. Cleavage of 32P-labeled R33B by the RNase H activity of both RTs was monitored with hybrids containing primers of 20 and 23–27 nucleotides. Reactions were carried out in the presence of 30 nm template-primer and 50 nm RT, and aliquots were withdrawn at 0.25, 0.5, 1, 5, 15, 30, and 60 min. Both enzymes were assayed in the absence and in the presence of dNTPs. In polymerization-dependent RNase H activity assays, the concentration of each nucleotide was 500 μm.