一种提取小鼠表皮总RNA的新方法

A New Method for Total RNA Isolation from Mouse Epidermis

目的:建立一种从小鼠表皮组织提取高质量RNA的方法。方法:用热击法分离小鼠表皮,用TRIzol法提取RNA,用紫外分光光度计测定RNA的产率和纯度,用琼脂糖电泳和RT-PCR检测RNA的质量和完整性。结果:采用新方法提取的小鼠表皮总RNA,其D260nm/D280nm值为1.8～2.0,大于1.5,且RNA产率高于100μg/g;琼脂糖电泳出现5S、18S和28S等3条清晰的rRNA条带,而且28SrRNA条带的亮度约为18S的2倍;用新方法制备的总RNA可成功地用于RT-PCR实验。结论:采用热击法分离表皮并结合TRIzol法可提取到高质量、完整性好的小鼠表皮总RNA,并能用于相关的分子生物学实验。

Objective： To construct a high performance method for isolating RNA from mouse epidermis. Methods： Mouse epidermis was separated after heating shock. Total RNA was extracted using TRIzol reagent. The amount and purity of isolated total RNA was detected by spectrophometer. RNA quality and integrality were detected by agarose electrophoresis and RT-PCR. Results： More than 100 lag total RNA could be obtained from 1 g mouse epidermal tissues. The absorbance ratios（D260nm/D280nm） of the total RNA ranged between 1.8 and 2.0 were over 1.5. After agarose electrophoresis, three bands representing 5S, 18S and 28S rRNA were clearly observed. The ratio of intensity of the 28S band to 18S rRNA band was about 2：1. Finally, RT-PCR was performed by using the isolated total epidermal RNA as templates. Conclusion： Total RNA with high quality and good integrality can be obtained from mouse epidermis by using TRIzol reagent after skin heating shock.