Figure 4
From: A mutant screening method by critical annealing temperature-PCR for site-directed mutagenesis
Verification of regression equation-derived T c by gradient PCR with different genes. As showed in the figure, the Tc identified by gradient PCR (hGH2V3-P233S: 70°C, pGHBP-S237C: 69°C and hGHR-R235C: 60°C) were very close to those derived from the regression equation (hGH2V3-P233S: TcF =69.21 and TcR =68.91; pGHBP-S237C: TcF = 68.54°C and TcR = 68.31°C; and hGHR-R235C: TcF = 59.81°C and TcR = 60.53°C). Although hGHR-R235C was inserted into pcDNA3.1(+) (5428 bp) differing from pET20b(+) (3716 bp), the Tc from PCR was almost same as that from the equation predicted.