一种适于枯草芽孢杆菌总RNA提取的改良方法

An improved method suitable for extraction of total RNA from Bacillus subtilis

枯草芽孢杆菌分泌多种胞外酶,采用普通试剂盒提取方法难获得完整、有效的RNA。在RNA试剂盒提取法的基础上,通过增加DEPC水洗涤菌体、溶菌酶裂解细胞、液氮冷冻预处理及调整RNA提取试剂添加比例等处理,可得到完整性良好的RNA。电泳检测结果表明,所获RNA条带清晰完整,无降解现象;提取的RNA可满足RT-PCR要求。

Bacillus subtilis can secrete lots of extra-cellular enzymes. Full total RNA is hardly extracted by the general total RNA kit method. On the base of the method of total RNA extraction kit, full total RNA may be extracted while washing cells in DEPC water, splitting cells by lysozyme, pretreating cells by liquid nitrogen and adjusting the volume proportion of the RNA extraction reagent. The electrophoresis results showed that RNA obtained was full without degradation. The RNA obtained could be used for RT-PCR.