[目的]优化仁用杏微卫星非变性聚丙烯酰胺凝胶电泳体系.[方法]以25份仁用杏和3份食用杏为材料,利用微卫星标记结合非变性聚丙烯酰胺凝胶电泳技术筛选出多态性高、可重复的SSR位点.并比较不同因子对电泳体系的影响,探索仁用杏微卫星非变性聚丙烯酰胺凝胶电泳的优化体系.[结果]仁用杏微卫星非变性聚丙烯酰胺凝胶电泳的优化体系为:聚丙烯酰胺凝胶的浓度为6%,丙烯酰胺与甲叉双丙烯酰胺的比例为29:1,1 000 V下电泳2～3 h,0.1% AgNO3 染色15 min 后显影.[结论]该优化体系为仁用杏资源微卫星标记遗传分析奠定技术基础.
Optimization of SSR-PCR Non-denatured Polyacrylamide Gel Electrophoresis Conditions in Kernelled Apricot
[Objective] The aim was to optimize the SSR-PCR non-denatured polyacrylamide gel electrophoresis conditions in kernelled apricot. [Method] 25 accessions of kernelled apricot and three accessions of edible apricot were selected as experimental materials to screen the repeatable SSR loci with high polymorphism by the use of SSR markers combined with non-denatured polyacrylamide gel electrophoresis. And the effect of different factors on electrophoresis conditions was compared to explore the optimal SSR-PCR non-denatured polyacrylamide gel electrophoresis conditions in kernelled apricot. [Result] The optimal non-denatured polyacrylamide gel electrophoresis conditions for SSR-PCR were established as follows: polyacrylamide gel concentration 6%, the ratio of acrylamide to bisacrylamide 29: 1, electrophoresis at 1 000 V for 2-3 h, and staining for 15 min within 0.1% AgNO3. [Conclusion] The optimum electrophoresis system has provided some technical foundations to further study the phylogenetic relationship of kernelled apricots by SSR markers.
Kernelled ApricotSSR markersPolyacrylamide gel electrophoresisSilver staining
NETL
NSTL
万方数据
维普
