Figure 4.

Effects of TthMutS on standard PCR amplification of mismatched templates. (A) Schematic representations of the primers and templates used in (B,C); (B) A perfectly matched primer was used to amplify perfectly matched (left), GT-mismatched (middle) or unpaired T-containing (right) templates; (C) Relative amounts of amplified fragments from reactions using perfectly matched (blue), GT-mismatched (red) or unpaired T-containing (purple) templates were plotted against the TthMutS concentration. The amounts of the products were normalized by those at 0 μM TthMutS; (D) The effect of TthMutS on amplification of the 80-bp mismatched template was examined by real-time PCR. The perfectly matched (blue), GT-mismatched (red) and unpaired T-containing (purple) templates were amplified by AmpliTaq Gold DNA polymerase by using perfectly matched primers. The threshold cycle (C_t) was determined and plotted against the TthMutS concentration. Experiments were repeated three times. Bars indicate standard deviations.