中文摘要：

高黏度发酵液具有菌体不易分离的特点。为了从γ-聚谷氨酸高黏发酵液中提取细菌总核糖核酸，利用磷酸缓冲液稀释发酵液、离心收集、焦碳酸二乙酯水洗涤以及溶菌酶处理的四步法收集菌体并制备核糖核酸待分离样品，进而分别采用RNAisoplus提取法，EZ-10RNA Miniprepkit试剂盒及PureLink RNA Minikit试剂盒法提取总核糖核酸，并通过琼脂糖凝胶电泳、核酸浓度检测仪和反转录聚合酶链式反应比较了三种方法提取总核糖核酸的效果。结果表明，四步法获得的菌体样品可以满足后续总核糖核酸提取的要求，三种方法提取的核糖核酸均能满足一般反转录聚合酶链式反应的要求，其中PureLink和RNAiso提取的核糖核酸A260nm／A230nm〉2．0，纯度较好，PureLink具有快速提取的优势而RNAiso则适用于大量样品的提取。

英文摘要：

The bacteria are difficult to be separated in the high viscosity fermentation broth. In order to extract bacterial total RNA from high viscosity fermentation broth ofpoly-γ-glutamic acid, four steps were taken to obtain the bacteria and prepare sample containing RNA used for separation. The four steps included dilute fermentation broth by phosphate buffer solution, centrifugation, wash cell by DEPC water and lysozyme treatment. Then, total RNA were extracted by RNAiso plus method, EZ-10 RNA Miniprep kit method and PureLink RNA Mini kit method. The RNA extraction effects of agarose gel electrophoresis, nucleic acid concentration detector and reverse transcription PCR were compared. Results showed that the sample obtained by the four-step method met the requirements of total RNA extraction. Total RNA extracted by the three methods all met the requirements of the RT-PCR experiments. However, the A260n m/A230nm of RNA that extracted by RNAiso plus method and PureLink RNA Mini kit method were larger than 2.0, and both methods shoned good purity. The PureLink Mini it method had the advantages of rapid extraction and the RNAiso was suitable for the extraction of large quantities of samples.