大白菜非变性聚丙烯酰胺凝胶电泳技术的优化  

The Optimization of Technique of Native Polyacrylamide Gel Electrophoresis(Native-PAGE)

作　　者：梁宝萍[1] 原玉香[2] 朴凤植[1] 张晓伟[2] 蒋武生[2] 梁爽[3] 姚秋菊[2] 张强[2] 赵艳艳[2] 

机构地区：[1]河南农业大学园艺学院,河南郑州450002 [2]河南省农业科学院园艺研究所,河南郑州450008 [3]郑州大学生物工程系,河南郑州450000

出　　处：《河南农业科学》2012年第5期129-132,136,共5页Journal of Henan Agricultural Sciences

基　　金：河南省创新杰出人才计划(094200510016);国家大宗蔬菜产业技术体系(CARS-25-G-27)

摘　　要：非变性聚丙烯酰胺凝胶电泳技术的检测效果受到凝胶聚合速度、片段迁移率等多种因素影响。以大白菜为供试材料,研究比较了凝胶聚合速度及PCR产物在不同浓度凝胶上的检测效果。不同浓度的凝胶是由2种〔40%(19∶1)和30%(29∶1)〕贮存液分别配制的4种浓度(5%,7%,9%,11%)。结果表明,在4～30℃,凝胶聚合速度随温度的升高而加快;APS保持不变,不同温度下TEMED∶APS合适配比也不同:4℃(冬季)时,TEMED/APS的合适配比是1∶10,25℃(春秋)、30℃(夏季),选择1∶20的配比为宜;2种不同贮存液配制的相同浓度凝胶检测效果不同,30%(29∶1)优于40%(19∶1),由前者配制7%的凝胶上DNA片段分离效果好,带型清晰,分辨率高。Many factors including gel＇s polymerization rate and fragments＇ migration ratio affect detection effect of native polyacrylamide gel electrophoresis（Native-PAGE）.The gel＇s polymerization rate and the detection effect of PCR products by two kinds of gel stock solution（40% Acr-Bis（19∶1） and 30% Acr-Bis（29∶1） prepared in four concentrations（5%,7%,9%,11%） were studied in this paper.The results showed that the gel＇s polymerization rate accelerated with increasing temperature in the range of 4-30 ℃,APS remains the same at different temperatures and TEMED∶APS suitable ratio was different： at 4 ℃（winter）,the ratio was 1∶10,at 25 ℃ or 30 ℃（spring and Autumn）（summer）,the ratio of 1∶20 was better.Detection of the effect was different in gel by two different stock solution prepared same gel concentrations;30% Acr-Bis（29∶1） is better than 40% Acr-Bis（19∶1）,and DNA fragment separation revealed clear band type,high resolution in the gel that was prepared using 7% stock solution.

关 键 词：非变性聚丙烯酰胺 聚合速度 贮存液 TEMED与APS配比 

分 类 号：Q784[生物学—分子生物学]