文冠果花药总RNA提取方法研究

以文冠果花药为材料 ,利用TRIZOL试剂快速提取法、异硫氰酸胍法和CTAB法提取花药总RNA .通过RNA产率、纯度、电泳图谱和mRNA差异显示等分析来确立适于文冠果花药RNA分离的方法 .研究结果表明 ,CTAB法提取的RNA呈现出 2 8SrRNA、18SrRNA和 5SrRNA三条较清晰的条带 ,很少有降解 ,其A2 6 0 A2 80 值可达 1 94 4 ,A2 6 0 A2 30 值为2 16 5 ,具有很高的纯度 .另 2种方法获得的RNA纯度较低 ,降解严重 ,有较多小分子和盐存在 ;RNA无明显条带出现 ,而是以小分子RNA弥散状分布 .使用差异显示法分析发现 ,用CTAB法提取的花药总RNA可得到理想的扩增条带 .

Method for RNA isolating from anthers of Xanthoceras sorbi-folia Bunge

The authors isolate total RNA from anthers of Xanthoceras sorbifolia Bunge by the methods of TRIZOL reagent, Guanidine Thiocyanate and CTAB and establish the method of RNA isolation suitable for anthers of Xanthoceras sorbifolia Bunge through the rate of yield, purity, electrophoresis and DDRT PCR of RNA. The result shows that there are three clear bands_28S rRNA, 18S rRNA and 5S rRNA respectively_and little RNA decomposes by the method of CTAB reagent, but RNAs isolated by the two other methods disperse in the pattern of small molecular instead of bands. And the purity of the former is higher than that of the latters, its ratio of A 260 /A 280 is 1.944 and the value of A 260 /A 230 is 2.165. RNA isolated by the methods of TRIZOL reagent and Guanidine Thiocyanate decompose seriously into small molecular, mixed with salt ion. Data of mRNA differential display show that there are ideal amplified bands using the RNA isolated by the method of CTAB reagent.