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Plant Diversity ›› 2008, Vol. 30 ›› Issue (05): 593-596.DOI: 10.3724 SP.J.1143.2008.07272

• 研究论文 • 上一篇    下一篇

一种从苏铁叶片中有效提取RNA 的方法

李 璐1 , 付乾堂1 , 2 , 余迪求1   

  1. 1 中国科学院西双版纳热带植物园, 云南昆明 650223 ; 2 中国科学院研究生院, 北京 100049
  • 收稿日期:2007-11-21 出版日期:2008-10-25 发布日期:2008-10-25

An Effective Protocol for the Isolation of RNA from Cycad Leaves

LI Lu1 , FU Qian-Tang1 , 2 , YU Di-Qiu1   

  1. 1 Xishuangbanna Tropical Botanical Garden , Chinese Academy of Sciences, Kunming 650223 , China;
    2 Graduate University of Chinese Academy of Sciences, Beijing 100049 , China
  • Received:2007-11-21 Online:2008-10-25 Published:2008-10-25

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摘要：

由于苏铁( Cycas revoluta) 叶片中含有大量的多糖多酚等次生代谢物, 常规RNA 提取方法很难获得优质的RNA。在常规的CTAB 法中加入了硼砂和β- 巯基乙醇来消除多酚和多糖的干扰, 得到了一个从苏铁叶片中有效提取RNA 的方法, 每克鲜叶片可获得约930μg RNA。A260 280 和A260 230 的纳米波长的吸收比值都约为2 , 表明RNA 的质量较好。获得的RNA 可用于Northern blot 和反转录PCR 等分析, 也说明RNA 的质量比较好。此外, 改进的提取方法也适合于含有次生代谢产物的其它植物, 同样可以获得优质RNA。

关键词: 苏铁, 硼砂, RNA 提取

Abstract: Conventional RNA extraction methods have been shown to produce poor-quality RNA when applied to Cycas revoluta because of abundant secondary metabolites included. With modification of the standard cetyltrimethylammonium bromide (CTAB) method by adding borax (disodium tetraborate decahydrate ) andβ-mercaptoethanol to eliminate the interference of polyphenol and polysaccharides , an effective protocol was developed . This modified protocol could extract high quantities of RNA from cycad leaves at about 930μg per gram of fresh weight . Both A260􊄯230 and A260􊄯280 ratios were around 2, indicating that RNA is of high quality. The RNA quality was confirmed by Northern blotting analysis and reverse transcription polymerase chain reaction (RT-PCR). The modified protocol could be successfully extended to other plants containing secondary metabolites .

Key words: Cycas revoluta

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