【摘要】 用修改了的异硫氰酸胍法从两种森林昆虫Clostera an-astomosis和Saperda populnea的不同组织中快速提取总RNA。电泳检测和cDNA合成分析结果表明,RNA未降解。经紫外分光光度计检测,A260/A280在1.8到2.0之间,表明RNA的纯度很高。用RT-PCR合成的双链cDNA的长度大于2kb,表明mRNA的完整性。用PCR的方法克隆了C.anasto-mosisβ肌动蛋白和几丁质酶的基因片段,表明RNA可用于其它分子操作。使用这个方法,在4个小时内可至少从8个样品中提取RNA并进行电泳分析。这些结果表明用这个修改后的一步法提取昆虫总RNA是省时,省钱和有效的。图3参10。更多还原

【Abstract】 A modified guanidinium isothiocyanate method was used to extract total RNA from two forest insect species Clostera anastomo-sis and Saperda populnea. The integrity of RNA was demonstrated by the methods of gel electrophoresis and cDNA analysis. Typical A260/ A280 absorbance ratio of the total RNA was in range of 1.8 to 2.0. The size of double strand cDNAs obtained by RT-PCR was more than 2 kb, which indicated that intact mRNA was obtained. The fragments of β-actin and chitinase gene from the RNA of C. anastomosis were ob-tained by RT-PCR, which indicated that the RNA could be used for other molecular operation. By this procedure, RNAs could be extracted and analyzed by electrophoresis from at least 8 samples within 4 hours. These results showed that this method was time- and cost-saving and effective.更多还原