›› 2009, Vol. 21 ›› Issue (6): 0-543.

• 论文 •    

EST-SSR荧光标记毛细管电泳检测法在豌豆上的应用及评价

龚亚明1,胡齐赞1,毛伟华2,李亚丹2,张古文1,丁 桔1   

  1. 1浙江省农业科学院 蔬菜研究所,浙江 杭州 310021;2浙江大学 分析测试中心,浙江 杭州 310029
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2009-11-25 发布日期:2009-11-25

Application and evaluation of fluorescent EST-SSR markers detection technique with capillary electrophoresis in pea

GONG Ya-ming;HU Qi-zan;MAO Wei-hua;LI Ya-dan;ZHANG Gu-wen;DING Ju   

  1. 1 Institute of Vegetables,Zhejiang Academy of Agricultural Sciences,Hangzhou 310021,China;2 Center of Analysis and Measurement,Zhejiang University,Hangzhou 310029,China
  • Received:1900-01-01 Revised:1900-01-01 Online:2009-11-25 Published:2009-11-25

摘要:

试验采用MegaBACE 1000 DNA 分析系统,利用荧光标记豌豆EST-SSR引物,建立豌豆EST-SSR荧光标记毛细管电泳检测法,并对该检测法的特点进行了分析和评价。结果表明,试验所建立的毛细管电泳荧光标记检测法能准确地识别片段大小、重复性好,可以明确区分不同等位基因;同时,通过采用两种不同颜色的荧光标记,可使检测192个样本的时间缩短在2 h之内。可见,本试验所建立的毛细管电泳荧光标记检测法具有高通量、高效率、高精确性等优点。


关键词: 豌豆, EST-SSR, 荧光标记, 电泳

Abstract: To detect EST-SSR markers in pea,a capillary electrophoresis with fluorescence detection was developed and evaluated based on fluorescence labeled EST-SSR primers and MegaBACE 1000 DNA analysis system. The result showed high precision and reproducibility in determining pea EST-SSR alleles by this detection system. Furthermore,the detection and analysis of 192 samples could be automatically completed within two hours by two different-color fluorescences labelled EST-SSR primers. Therefore,it is suggested that the capillary electrophoresis with fluorescence detection developed in our study with high throughput,efficiency.

Key words: pea(Pisum sativum L.), EST-SSR, fluorescent marker, electrophoresis