表皮葡萄球菌总RNA小量提取方法比较

Comparison of methods for total RNA extraction from Staphylococcus epidermidis

表皮葡萄球菌属于革兰氏阳性细菌,因细胞壁较厚、难破裂等特点致使其总RNA提取的难度增大,而总RNA提取效率和提取质量等对分子生物学相关研究起着关键的作用。因此,以表皮葡萄球菌ATCC35984为实验菌株,分别使用Trizol裂解联用研磨珠破碎法、单酶酶解法、双酶酶解法、液氮研磨法等4种细胞壁破碎方法,对其总RNA进行提取,对比提取效果,旨在寻找一种高效稳定的总RNA提取方法。结果显示,双酶裂解法和液氮研磨法提取的表皮葡萄球菌总RNA完整性好、纯度高、重复性好,可有效提高表皮葡萄球菌总RNA提取质量。

The efficiency and quality of extracted total RNA play a key role in molecular biology research.Staphylococcus epidermidis,a gram positive bacterium,possesses thick cell wall,which is hard to rupture and difficult to extract its total RNA.In this study,different strategies including single enzyme disruption,combined enzyme disruption etc.were compared in order to obtain an efficient and stable method for total RNA extraction from S.epidermidis ATCC35984.Results showed that the total RNA extracted by liquid nitrogen grinding or combined enzyme disruption exhibited relatively better integrity,purity and repeatability.