改良TRIzol法高效提取东北林蛙皮肤总RNA

An Improved TRIzol Method to Extract Total RNA from Skin Tissue of Rana dybowskii

目的:建立一种从两栖动物皮肤组织中提取高质量总RNA的方法。方法:改良传统TRIzol法,提取皮肤总RNA,紫外分光光度计测定RNA纯度和得率,1.1%琼脂糖凝胶电泳检测其完整性。结果:改良方法提取的总RNA,其A260/A280值在1.8～2.1之间,A260/A230大于2;琼脂糖凝胶电泳显示清晰的28s rRNA和18s rRNA条带,且28s rRNA条带亮度约为18s rRNA的2倍。结论:改良TRIzol法提取的总RNA纯度高、完整性好、杂质少、得率大,易于操作掌握,可以用于相应的分子生物学试验。

Our objective was to develop a method for extracting high quality total RNA from amphibian skin tissue.We extracted total RNA from the skin of Dybowski＇s frog using a modified traditional TRIzol method.We assayed the purity and yield of the RNA by UV spectrophotometer.We examined the integrity of the extracted RNA by 1.1%agarose gel electrophoresis.The A260/A280 ratios of RNA extracted by our modified method ranged from 1.8 to 2.1,and the A260/A230 ratios were greater than 2.Agarose gel electrophoresis showed clear 28s rRNA and 18s rRNA bands,and the brightness of 28s rRNA bands was approximately twice the 18s rRNA.We conclude that the purity and integrity of total RNA extracted by our improved TRIzol method were higher than that extracted by the traditional TRIzol method：the total RNA had fewer impurities and the yields were larger.The modified method was easier to implement and can be used in relevant molecular biology tests.