[目的]改良SSR标记变性聚丙烯酰胺凝胶电泳(PAGE)技术,提高大规模SSR分子标记分析的工作效率.[方法]在利用SSR标记构建中国春/兰考大粒F2群体遗传图谱的基础上,对变性PAGE点样方法进行改进,并对中国春/兰考大粒筛选的122对差异引物进行组合分析.[结果]试验表明,引物扩增条带特异性强、覆盖范围不大即可采用两次点样法分析；两次点样和单次点样的凝胶扩增条带均清晰可辨;分析相同数量的样品,两次点样法较单次点样法节约了41％的试验时间,节约了50％的试剂用量.[结论]两次点样法简化了试验步骤,节约了试验成本,加速了试验进程,更适用于遗传作图.%[Objective] To improve the efficiency of genetic analysis and the methods for spotting of denatured PAGE in detecting SSR. [ Method] The technique of denatured PAGE in detecting SSR is extremely sensitive electrophoretic separation. Due to the over many technical steps and large time consumption, this technique was disadvantageous to carry out molecular marker analysis in large scale. This research improved the methods for spotting, at the base of the construction of the genetic linkage maps for F2 from the cross between Chinese spring and Lankaodali using SSR markers. [ Result] The results were as following: Screening of Chinese spring and Lankaodali, total of 122 SSR markers were different. As long as bands of high specificity, the markers could be detected by twice spotting. The bands by twice spotting and once spotting were both distinguished, the same number of samples were analyzed, the method of twice spotting saved 41% time and 50% reagents. [ Conclusion] The method of twice spotting simplified experimental procedures, saved experimental cost, accelerated test process and was suitable for genetic mapping.
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