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大白菜SSR检测体系的优化

文献类型: 中文期刊

作者: 石磊 1 ; 李丽 2 ; 郑晓鹰 2 ;

作者机构: 1.甘肃农业大学草业学院

2.北京市农林科学院蔬菜研究中心

关键词: 大白菜;SSR;检测体系;优化;SDS-聚丙烯酰胺凝胶电泳

期刊名称: 分子植物育种

ISSN: 1672-416X

年卷期: 2007 年 5 卷 01 期

页码: 110-116

收录情况: CSCD

摘要: 以不同地区栽培的5份大白菜品种为试材,从PCR反应组成、扩增程序、电泳检测等环节对SSR技术进行了优化,建立了一套适用于大白菜品种鉴定的SSR-PCR体系。即10μL反应组成为:1×buffer;2.50mmol/LMgCl2;0.10mmol/LdNTPs;0.35μmol/LSSR引物;1.00ng/μL模板DNA和0.40UTaq酶。适宜的扩增程序为72℃热启动3min,94℃预变性2min后进行20个迫降循环:94℃变性60s,68℃退火60s,72℃延伸45s(-1℃/2cycles);再进行20个循环:94℃变性60s,58℃退火60s,72℃延伸45s,72℃延伸10min。应用SDS-聚丙烯酰胺凝胶(银染)电泳检测并取得很好的效果。选用108对芸薹属SSR引物,对这5份品种的基因组进行扩增,筛选出具有2条以上特异扩增条带的SSR引物22对。用这些引物对5份品种进行扩增,初步分析了SSR标记的多态性及用于大白菜指纹分析的潜力。

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