The application of asymmetric PCR-SSCP in gene mutation detecting

Abstract

The advantages and disadvantages of asymmetric PCR-SSCP and the traditional PCR-SSCP were compared in this study. The mutations in 3′UTR of Smad4 gene of Luxi cattle and the Holstein cow were analyzed by asymmetric PCR-SSCP and one insert “T” mutation and one G/A mutation in this region were found. The G/A mutation created a HhaI restriction enzyme digestion position and the frequencies studied by asymmetric PCR-SSCP and HhaI-RFLP in 116 Luxi cattle and 75 Holstein cows were all the same. The asymmetric PCR-SSCP had fewer, clearer and more stabile bands than traditional PCR-SSCP. This indicates that the asymmetric PCR-SSCP is suited for mutation detection.

Keywords

asymmetric PCR-SSCP / PCR-RFLP / mutation / Smad4