FIGURE 5.
Real-time fluorescence-based analysis of RNA digestion by the 3′–5′ exoribonuclease activity of RNase PH. (A) RNA2 was designed to be sensitive to potential 3′–5′ exoribonucleases. Based on the same procedure mentioned in Figure 1, RNA2–DNA2 duplex formation brings the TAMRA into the vicinity of FAM and to quench FAM fluorescence emission. (B) In the presence of RNase PH exoribonuclease, the increase in FAM fluorescence signal can be analyzed in real time. RNA2 and DNA2 concentrations are, respectively, 0.5 μM and 1 μM.
