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农学学报

农学学报 ›› 2012, Vol. 2 ›› Issue (7): 25-28.

所属专题： 园艺

• 农艺科学 作物遗传育种 生理生化 • 上一篇    下一篇

核桃芽总RNA提取及RT-PCR

宋艳波 梅霞 吴国良 牛洪斌   

  • 收稿日期:2012-06-07 修回日期:2012-06-14 出版日期:2012-07-20 发布日期:2012-07-20
  • 基金资助:

    桃果实韧皮部糖卸载及转运蛋白基因克隆及表达特征

Total RNA Extraction and RT-PCR of Walnut Bud

  • Received:2012-06-07 Revised:2012-06-14 Online:2012-07-20 Published:2012-07-20

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1392

摘要：

为了从核桃芽体内提取高质量的RNA，以便进行核桃成花基因克隆研究，针对核桃芽体内富含多酚等次生代谢物的特点，在总结他人研究基础上对SDS法和CTAB法及试剂盒提取法进行了改良和比较。凝胶电泳检测表明，Spectrum Plant Total RNA试剂盒提取法及以pH 9.0硼砂为缓冲液的改良CTAB法和改良SDS法所提取RNA的28SrRNA和18SrRNA条带清晰，且28SrRNA的亮度约是18SrRNA的2倍，说明所提RNA完整性较好；紫外光谱分析显示，3种方法的OD260/OD280分别为1.99、1.96、1.77，OD260/OD230均大于2.0，RNA产率依次为360、180和75 μg/(g?FW)。综合检测结果认为，改良CTAB法和改良SDS法所提RNA的完整性和纯度均能满足基因克隆等的研究，是一种适合核桃芽体内RNA提取的经济而有效的方法。

关键词: 莫兰蒂台风, 莫兰蒂台风, 行道树, 对策, 厦门市

Abstract:

Extracting high-quality total RNA from walnut buds is a complicated and rigorous precess as the walnut tissue is rich in phenolic compounds、polysaccharides and other secondary metabolites, this may influence the follow-up molecular biology research. In this paper, we discussed different extraction method of Walnut buds and obtained a effective method. The results showed that added PVPP to the grinding and the use of modified SDS method of PH9.0 sodium borate buffer and the modified CTAB method can extract walnut buds RNA well,and we got the target fragment with the reaction of RT-PCR.So we can acquire the conclusion that our method about extracting walnut bud RNA is available to be used for the research ahout Walnut’s flowering gene .

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