PCR-SSCP检测微卫星基因型方法的研究

Application of PCR-SSCP in genotyping microsatellite loci

非变性丙烯酰胺凝胶电泳(Native-PAGE)在检测片段差异较小的微卫星等位基因时误差较大。针对Native-PAGE的不足,以鸡1号染色体上的7个微卫星位点为例,对PCR-SSCP(单链构陷多态)检测此类等位基因的效果进行了分析。结果表明,对MS202和MS502,通过检测单链DNA,PCR-SSCP能准确分离PAGE无法区分的片段差异较小的等位基因;对MS1404,单链和双链DNA分离效果相同;对MS1805、MS1803、wxt_MS_3,双链DNA得到了很好分离,但单链DNA分离效果不佳。结果表明,对某些微卫星位点,PCR-SSCP能弥补Native-PAGE不足。

Native polyacrylamide gel electrophoresis （PAGE） is not powerful in separating microsatellite alleles with almost same length of DNA fragments. To complement the deficiency of PAGE, we attempted to detect genotype of seven microsatellite loci located on chicken chromosome 1 using single strand conformation polymorphism （SSCP）. Results showed that SSCP could clearly separate alleles with almost same size in MS202 and MS502, while it was very difficult for PAGE to achieve the result. Both double and single strand DNAs were successfully separated in MS1404. In MS1805, MS1803 and wxt_MS_3, double strand DNA was successfully separated, but not for single strand DNA. The data indicate that SSCP can complement deficiency of PAGE in separating alleles with almost same size in some microsatellite loci.