提取猕猴桃果实总RNA的新方法

目的:获得1种提取猕猴桃果实总RNA的简单、快速、高效的方法。方法:使用异硫氰酸胍提取剂,同时在提取剂中加入1/10体积的5mol/L醋酸钾及1/4体积的100%冰乙醇提取猕猴桃果实总RNA。将经检测合格的RNA反转录为cDNA。以上述cDNA为模板,通过PCR方法扩增猕猴桃果胶甲基酯酶抑制剂基因。结果:所提取的猕猴桃果实总RNA可完全消除果实组织多糖的干扰,并在实验室成功地用于cDNA的合成和PCR扩增反应等。结论:与其它提取方法相比,本方法具有操作步骤简单、耗时少等优点,还可用于其它富含多糖的果实组织(如番茄等)的RNA提取。

A New Method for Extracting Intact RNA from Kiwi Fruit

Objective:To acquire a rapid and simple procedure for efficiently extracting total RNA from kiwi fruit. Method:Using the guanidinium thiocyanate extraction buffer,together with adding 1/10 volume of 5 mol/L potassium acetate and 1/4 volume of 100% ice ethanol,total RNA was extracted from kiwi fruit. The good quality RNA was reverse transcribed into cDNA,and the synthesized cDNA was used as the template for PCR amplification of kiwi pectin methylesterase inhibitor. Results:This method could entirely eliminate interference of polysaccharides. The RNA isolated by this method had been successfully used in our laboratory for cDNA transformation and PCR amplification,and so on. Conclusion:Comparing with other methods,the operational steps of this method were very simple and the consumed time was also little. In addition,the method was able to be used efficiently for the extraction of intact RNA from other fruit tissues rich in polysaccharides,such as tomato.