大白菜非变性聚丙烯酰胺凝胶电泳(PAGE)技术影响因素研究

Research on Influence Factor of Non Denatured PAGE Technology of Chinese Cabbage

电泳技术是分子标记辅助育种技术的关键步骤,建立一套高效的非变性聚丙烯酰胺凝胶电泳技术体系可提高分子标记辅助育种技术的工作效率。以根肿病抗感大白菜作为试材,以凝胶聚合速度及凝胶条带的清晰度作为衡量标准,分别对电泳中的凝胶浓度、温度、促凝剂配比3个重要因素进行研究。结果发现凝胶浓度、温度、促凝剂配比均与凝胶聚合时间呈现负相关,但4种常用凝胶浓度制胶时间差异不明显,可主要根据目的产物的大小选择浓度。在凝胶质量方面,温度、促凝剂配比与凝胶的脆性呈负相关。综合分析,操作中环境温度为25~35℃,促凝剂配比为1∶10的条件,聚合速度适中,条带清晰,检测效果最佳。

The electrophoresis technology is the key step in the molecular marker assisted breeding technology.Establishing a set of effective system of modified polyacrylamide gel electrophoresis technology could improve the working efficiency of the molecular marker assisted breeding technology.Taking Chinese cabbage with resistance to club root as test materials,polymerization time to gel and the definition of the gel strips as standards,the concentrations of the gel temperature and ratio of coagulant were studied.Found that the gel concentration,temperature,ratio of coagulant were negatively correlated with gel time,but the differences of gel time by four kinds of commonly used gel concentration were not obvious,we can choose the concentration mainly according to the size of the objective.In terms of gel quality,the temperature and the ratio of coagulant were negatively correlated with the brittleness of the gel.Therefore,comprehensive analysis showed that the polymerization speed was moderate,the strip was clear,and the detection effect was the best when the environmental temperature was 25-35℃,the ratio of coagulant promoter was 1∶10.