四种副溶血弧菌总RNA提取方法的比较

Comparison of four Kinds of Methods for Extracting Total RNA from Vibrio parahaemolyticus

副溶血弧菌(Vibrio parahaemolyticus)是一种革兰氏阴性嗜盐菌。提取高质量的RNA是研究副溶血弧毒力基因表达与其致病性和环境适应性的基础。本研究分别用SDS法、Trizol法,PureLinkTM RNA Mini Kit和Uniq-10柱式Trizol总RNA抽提试剂盒提取六株副溶血弧菌的总RNA,用普通琼脂糖凝胶电泳和核酸测定仪检测RNA的质量,并用RT-PCR法对四种提取方法进行比较。研究结果表明,Trizol法和Pure-LinkTM RNA Mini Kit简单快捷,提取的总RNA完整性好,纯度高,可用于后续实验的分子生物学研究;而Trizol法更适用于普通实验室细菌RNA的提取。

Vibrio parahaemolyticus is a kind of halophilic gram-positive bacterias.Extracting high-quality total RNA from Vibrio parahae molyticus is essential for studying virulence gene expression and its pathogenicity and environmental adaptability.In this research,we extracted total RNA from six strains by using SDS method,Trizol method,PureLinkTM RNA Mini Kit and Uniq-10 column Trizol total RNA extraction kit.Then detected the quality of extracted RNA with ordinary agarose gel electro phoresis and nucleic acid detector,and comparison of four kinds of extraction methods by using RT-PCR method.The results showed that Trizol method and PureLinkTM RNA Mini Kit are simple and rapid,which total RNA isolated byabove two methods has the characteristic of good integrity and high purity,and can be used for subsequent molecular biology research.However,Trizol method approriated for isolating total RNA of bacterial from ordinary laboratory.