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中国农学通报

中国农学通报 ›› 2010, Vol. 26 ›› Issue (2): 152-156.

• 林学 园艺 园林 • 上一篇    下一篇

杏叶片与果实总RNA提取方法研究

李晓颖,曹雪,房经贵,章镇   

  • 收稿日期:2009-09-02 修回日期:2009-09-24 出版日期:2010-01-20 发布日期:2010-01-20

Study on Methods for RNA Extraction from Apricot Leaf and Fruit

  • Received:2009-09-02 Revised:2009-09-24 Online:2010-01-20 Published:2010-01-20

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摘要：

以杏叶片及幼果为材料，分别从操作耗时、RNA质量和产量等方面比较了CTAB法、改进CTAB法、SDS法、改进SDS-酚法和RNA提取试剂盒Trizol等5种不同的RNA提取方法。结果表明：5种方法均能从幼嫩叶片中提取到总RNA；经电泳检测，28SrRNA和18SrRNA两条主带清晰，A260/A280大于1.8。在叶片RNA提取过程中，改良的CTAB法及Trizol法能获得高质量的RNA，产量也分别达到395.21和839.25μg/g。其中改良SDS以及改良CTAB能更有效减少果实RNA的提取中酚类物质和多糖等杂质的影响，获得质量高的总RNA。每种方法提取的叶片和果实的RNA经反转录后形成cDNA，并进行RT-PCR扩增，成功扩出看家基因泛素蛋白UBQ片段，说明了RNA能够很好地应用于文库构建、基因克隆等研究。

关键词: 铜, 铜, 人工瘤胃, VFA, 瘤胃发酵

Abstract:

The efficiency and the effect of five RNA isolation methods, including CTAB, SDS, modified CTAB, modified SDS and Trizol, were studied in the RNA extraction from apricot leaves and fruitlets. Of these five methods, the method of Trizol was found to be the best one, and the modified CTAB was better for the leaf RNA isolation. The RNA yields from apricot leaf were 839.25μg/g and 395.21μg/g, respectively. The modified CTAB and SDS methods were preferential ones for the isolation of higher quantity and better quality RNA from apricot fruit. All the RNA samples could be successfully used to amplify a specific cDNA fragment of UBQ, a housekeeping gene in plant, which suggests the utility of the RNA samples in cDNA library construction or gene cloning.