Simultaneous Use of MutS and RecA for Suppression of Nonspecific Amplification during PCR
Figure 1
The error-suppressing effects of ttRecA and ttMutS in the presence of ATP. (a) A schematic representation for the mechanism by which ttMutS suppresses nonspecific amplifications during PCR. A ttMutS dimer recognizes mismatched bases generated by mishybridization of the primer and blocks the approach of DNA polymerase. (b) A schematic representation for the mechanism by which ttRecA suppresses nonspecific amplification during PCR. ttRecA promotes proper priming for PCR. (c) A 423 bp region of the ttha1806 gene was amplified by using Takara LA Taq in the presence of 0 to 0.4 mM ATP. Lanes 1–4, 5–8, and 9–12 are the results of the reaction without ttMutS or ttRecA, with 0.8 M ttMutS, and with 0.4 M ttRecA, respectively. The amounts of the amplified fragments were quantified by using the ImageJ software [9] and are shown as bar graphs in the lower panels, where gray and blue indicate nonspecific and desired amplifications, respectively. (d) A 423 bp region of the ttha1806 gene was amplified by using Takara LA Taq in the presence of 0.9, 2.7, 8.0, or 24 ng/mL template DNA (T. thermophilus HB8 genomic DNA). The relative amounts of the amplified fragments are shown. Gray and blue bars indicate nonspecific and desired amplifications, respectively.
