变性与非变性PAGE在检测家蚕微卫星PCR产物中差异

The Difference in the Detection of SSR Products with Denatured and non-denatured PAGE

利用公布的家蚕基因组序列,设计特定的家蚕微卫星引物,对8个不同品种家蚕的微卫星位点进行PCR扩增,所得PCR产物经变性与非变性聚丙烯凝胶(PAGE)分离,银染后发现在非变性PAGE上,条带较粗且模糊,非特异带较多,导致读带误差增大;而在变性PAGE上,所得条带较细且比较清晰,非特异带明显减少,有利于降低统计误差。

Silkworm SSR primers were designed based on the silkworm genome sequence. SSR products were amplified from 8 silkworm varieties, and detected by denatured and non-denatured gels respectively. In non-denatured gel, there were many nonspecific bands and shadow bands, leading to misread of amplication. In denatured gel, nonspecific and shadow bands were greatly lessened while clear purposed bands were showed, in favor of decreasing the statistical error.