玉米微卫星PCR产物变性与非变性PAGE-银染法的比较分析

Comparison of Microsatellite Products of Maize Using Denatured and Non-denatured PAGE-Silver-Staining Methods

对变性聚丙烯酰胺凝胶电泳和非变性聚丙烯酰胺凝胶电泳进行比较,探讨建立简便、快速、安全、可靠、经济、效率高、毒害低的应用于玉米遗传多样性分析的SSR标记方法。结果表明,用2对SSR引物对6份玉米自交系基因组DNA进行扩增,将扩增产物在变性与非变性聚丙烯酰胺凝胶(PAGE)上电泳,银染后微卫星扩增产物在二者上的电泳结果存在差异。在变性胶中微卫星扩增产物目的条带清晰,易于鉴定;在非变性凝胶中表现有较多的非特异性条带,但目的条带很明亮,容易看出来,不影响实验结果。非变性聚丙烯酰胺凝胶电泳带型清晰准确、结果可靠、经济快速,能真实地揭示玉米自交系间的遗传多样性,是玉米种质类群划分的有效分子标记方法。

Comparing the denatured PAGE and non-denatured PAGE to establish a simple, rapid, secure, reliable, economic, high efficient and low toxic method of SSR molecular marker, which was applied to genetic diversity analysis of maize. Two microsatellite primer pairs were used to amplify the genomic DNAs of maize. The PCR products were detected by denatured and non-denatured polyacrylamide gel electrophoresis（PAGE）. The results of sil- ver-staining indicated that there were many distinct differences about products between the two kinds of gel. The tar- / geted bands clearly appeared in denatured gels, which were easy to identify. There were many nonspecific bands in non-denatured gels. However, targeted bands were very bright, easy to see and did＇t affect experimental results. The non-denatured gel could reveal genetic diversity analysis among maize inbred lines truly, which was an effective molecular marker to divide germplasm groups of maize.