2种提取大鼠肝RNA方法的比较

Comparison of two methods of extracting RNA from liver of rats

目的建立大鼠肝RNA的提取方法,为逆转录酶聚合酶链式反应(RT-PCR)、Northern blot以及cDNA文库构建等分子生物学实验奠定基础。方法用Trizol法和异硫氰酸胍提取法提取大鼠肝RNA,紫外分光光度计测定RNA产量及纯度,琼脂糖凝胶电泳验证RNA的完整性。结果采用Trizol方法提取的RNA的OD260/OD280比值为2.28,异硫氰酸胍方法提取的RNA OD260/OD280比值为1.74。采用Trizol法提取RNA的产量高于异硫氰酸胍提取法,电泳分析18 s、28 s条带完整。结论Trizol提取的大鼠肝RNA纯度、收率都优于硫氰酸胍方法提取的RNA。

Objective To establish the method of extracting total RNA from liver of rats for using in the reverse transcription polymerase chain reaction （RT - PCR）, Northern blotting and eDNA library construction further. Methods Two methods （Tfizol and guanidinium isothioeyanate） of extracting RNA from Rats liver were carried out, optieal density was measured under ultraviolet speetrophotometer,and the integrality of RNA were identified through agarose gel eleetrophoresis, Results OD260/ OD280 of RNA which was extracted through Trizol and guanidinium isothioeyanate methods was 2, 28 and 1.74 respectively. The purification quality of RNA extracted by Trizol was better than guanidium thioeyanate, and the 28S and 18S RNA of total RNA were intact. Conclusion The purity and yid of RNA extracted by Trizol was better than guanidium thiocyanate.