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毛细管与聚丙烯酰胺凝胶电泳检测甜菜SSR位点的比较研究
引用本文:冯琬淇,张福顺,刘乃新.毛细管与聚丙烯酰胺凝胶电泳检测甜菜SSR位点的比较研究[J].中国农学通报,2022,38(36):34-41.
作者姓名:冯琬淇  张福顺  刘乃新
作者单位:黑龙江大学现代农业与生态环境学院,哈尔滨 150080
基金项目:财政部和农业农村部国家现代农业产业技术体系(糖料)建设项目“质量安全与营养品质评价”(CARS-170503)
摘    要:为了探究毛细管与聚丙烯酰胺电泳方法的适用情况,比较其优缺点。本研究以8个甜菜栽培种资源为材料,选择8个SSR位点,分别使用聚丙烯酰胺凝胶电泳银染检测和毛细管电泳荧光检测每个位点的多态性,并对这两种方法的检测结果进行了比较。毛细管电泳荧光标记法可以准确的知道DNA片段的分子大小;PAGE电泳只能用肉眼与Marker比较估测得出大致分子量,不能得出准确分子量。8对引物两种检测方法的匹配比率分别是75%、87.5%、50%、75%、25%、100%、37.5%、50%。研究结果证明,两种检测平台数据整合存在较大差异。检测结果与引物是否是单拷贝、引物多态性,稳定性等因素有直接关系。测序检验结果表明,样品较少时利用PAGE电泳的方法来检测SSR位点的多态性,既能节省成本又能节省时间,还能保证检测结果的准确性。

关 键 词:甜菜  简单序列重复  聚丙烯酰胺凝胶电泳  毛细管电泳  DNA片段  
收稿时间:2022-06-19

SSR Polymorphism in Sugar Beet: Detection by Polyacrylamide Gel Electrophoresis and Capillary Electrophoresis
FENG Wanqi,ZHANG Fushun,LIU Naixin.SSR Polymorphism in Sugar Beet: Detection by Polyacrylamide Gel Electrophoresis and Capillary Electrophoresis[J].Chinese Agricultural Science Bulletin,2022,38(36):34-41.
Authors:FENG Wanqi  ZHANG Fushun  LIU Naixin
Institution:College of Modern Agriculture and Ecological Environment, Heilongjiang University, Harbin 150080
Abstract:In order to explore the applicability of capillary electrophoresis and polyacrylamide electrophoresis and compare their advantages and disadvantages, 8 sugar beet cultivation resources were used as materials, 8 SSR loci were selected. The polymorphism of each locus was detected by polyacrylamide gel electrophoresis silver staining and capillary electrophoresis. And then the results of the two methods were compared. Capillary electrophoresis fluorescence labeling method could accurately obtain the molecular size of DNA fragments. PAGE electrophoresis could only estimate the approximate molecular weight with the naked eye compared with Marker, but could not obtain the accurate molecular weight. The matching ratios of the two detection methods with the 8 pairs of primers were 75%, 87.5%, 50%, 75%, 25%, 100%, 37.5% and 50%, respectively. According to the results, there were great differences in data integration between the two detection platforms. The detection results were directly related to the factors like whether the primers were single copy, primer polymorphism and stability. The sequencing test results showed that when there were fewer samples, using PAGE electrophoresis to detect the polymorphism of SSR loci could not only save cost and time, but also ensure the accuracy of the test results.
Keywords:sugar beet  SSR  polyacrylamide gel electrophoresis  capillary electrophoresis  DNA fragments  
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