一种有效的可口革囊星虫体壁总RNA提取方法

[目的]为从可口革囊星虫体壁中提取高质量的RNA,以便深入开展分子生物学研究。[方法]采用Trizol法、改良Trizol法和十六烷基三甲基溴化铵(CTAB)法3种方法提取可口革囊星虫体壁中总RNA,并通过琼脂糖凝胶电泳比较不同方法提取总RNA的效果。[结果]改良Trizol法提取的总RNA经电泳后能看到清晰完整的28S rRNA、18S rRNA和5S rRNA条带,无拖尾现象；而Trizol法和CTAB法提取的总RNA电泳条带完整性较差,缺少28S rRNA。[结论]传统的Trizol法和CTAB法不适用于可口革囊星虫体壁总RNA的提取；改良Trizol法提取的总RNA电泳条带清晰、完整性好,质量符合cDNA合成、RT-PCR扩增等后续实验要求,适用于提取可口革囊星虫体壁总RNA。

Total RNA of the Body Wall of Phasolosma esculenta: An Effective Extraction Method

The paper aims to extract high quality RNA from the body wall of Phasolosma esculenta for further molecular biology researches. We extracted total RNA from the body wall of Phasolosma esculenta by the modified Trizol extraction method, the Trizol reagent method and the hexadecyl- trimethyl- ammonium- bromide (CTAB) method, and compared the effects of different methods on total RNA extraction by agarose gel electrophoresis. The results showed that: after the agarose gel electrophoresis of the total RNA extracted by the modified Trizol extraction method, we could see clear and complete 28S rRNA, 18S rRNA and 5S rRNA bands with no smear; however, the total RNA electrophoresis bands extracted by the Trizol reagent method and the CTAB method had poor integrity and lacked 28S rRNA. The Trizol reagent method and the CTAB method are not suitable for the extraction of total RNA from the body wall of the Phasolosma esculenta. The electrophoresis bands of the total RNA extracted by the modified Trizol extraction method are clear and have a good integrity, their quality meet the requirements of cDNA synthesis, RT- PCR amplification and other follow- up experiments.