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中国农学通报 ›› 2012, Vol. 28 ›› Issue (22): 34-38.doi: 10.11924/j.issn.1000-6850.2012-0803

所属专题: 园艺

• 林学 园艺 园林 • 上一篇    下一篇

板栗子叶总RNA提取方法研究?

王滨蔚 车鹏燕 何承忠 王猛 石卓功   

  • 收稿日期:2012-03-09 修回日期:2012-04-13 出版日期:2012-08-05 发布日期:2012-08-05
  • 基金资助:

    国家自然科学基金项目“中国板栗花粉直感效应及其机理研究”

Study on Methods of Total RNA Extraction from Cotyledon of Chinese Chestnut

  • Received:2012-03-09 Revised:2012-04-13 Online:2012-08-05 Published:2012-08-05

摘要:

为了获得板栗子叶总RNA提取的理想方法,为后续的分子生物学研究打下基础,以板栗幼果子叶为材料,采用改良CTAB法、硼砂-CTAB法、酚-SDS法、皂土法和2种植物总RNA提取试剂盒共6种方法提取板栗子叶总RNA。结果表明,除硼砂-CTAB法外,其他5种方法均能获得板栗子叶总RNA。但是,与酚-SDS法、皂土法和2种植物总RNA提取试剂盒相比较,改良CTAB法提取的板栗子叶总RNA完整性好,纯度高,且无DNA污染,能够满足cDNA-AFLP等分子生物学研究的要求。

关键词: 特征, 特征

Abstract:

In order to obtain an ideal total RNA extraction method of cotyledon in Chinese chestnuts for providing foundation for molecular biology research, total RNA of cotyledon excised from young fruit of Chinese chestnut were extracted with 6 methods, including improved CTAB method, borax-CTAB method, phenol-SDS method, bentonite method and 2 RNA extraction kits. The results showed that total RNA was obtained with 5 methods but borax-CTAB method. However, the improved CTAB method was better than phenol-SDS method, bentonite method and two RNA extraction kits. The total RNA extracted with improved CTAB method had high integrity, high purity and no DNA pollution, and would be suitable for related molecular researches, such as cDNA-AFLP markers.