Abstract：Abstract: Comparing with their corresponding wild allele genes, a single nucleotide mutation had been taken place in high pigment genes hp1 and hp2, respectively, which was very difficult to be discriminated by normal PCR technique. In this experiment, the relationship between specific PCR and mismatched bases as well as their positions were studied. Results showed that the primer with double-base mismatch TA-TG can distinguish mutation location of hp1 from its wild allele location at annealing temperature 49.3 +0.1 ℃. A primer with T-C mismatch can distinguish mutation location of hp2 from its wild allele location at annealing temperature 53.5+0.1 ℃. The postion of mismatched bases at 3` end of primers had little effect on specificity of PCR.
