《中国畜牧兽医》 ›› 2016, Vol. 43 ›› Issue (10): 2792-2796.doi: 10.16431/j.cnki.1671-7236.2016.10.042

• 经验交流 • 上一篇    下一篇

微波加热快速提取芽孢杆菌与乳酸菌DNA

周晏阳, 邱莞思, 唐俊妮, 汤承   

  1. 西南民族大学生命科学与技术学院, 成都 610041
  • 收稿日期:2016-03-29 出版日期:2016-10-20 发布日期:2016-10-28
  • 通讯作者: 汤承 E-mail:tangcheng101@163.com
  • 作者简介:周晏阳(1992-),男,黑龙江萝北人,硕士,研究方向:动物微生物学,E-mail:378292703@qq.com
  • 基金资助:

    “十二五”科技支撑计划(2012BAD13B06)

Rapid Extraction of Bacillus and Lactobacillus DNA by Microwave Heating

ZHOU Yan-yang, QIU Wan-si, TANG Jun-ni, TANG Cheng   

  1. College of Life Science and Technology, Southwest University for Nationalities, Chengdu 610041, China
  • Received:2016-03-29 Online:2016-10-20 Published:2016-10-28

摘要:

试验旨在研究快速、简单、高效提取芽孢杆菌与乳酸菌DNA的方法。在微波炉额定功率800 W下,对芽孢杆菌和乳酸菌进行微波处理40~150 s,离心获取上清,收获细菌DNA,将样本DNA PCR扩增后进行琼脂糖凝胶电泳验证,测定其纯度和产量后测序。结果显示,在微波炉功率800 W条件下加热40~150 s均可有效提取芽孢杆菌与乳酸菌DNA。所得DNA质量较好(D260 nm/D280 nm在1.8~2.1之间)。DNA浓度符合PCR检测要求,目的条带清晰,所得测序结果满足常规分子生物学研究。该微波提取方法具有简单、快速、高效的特点,且具有广泛的实用性,为乳酸菌与芽孢杆菌快速分子检测提供了简便手段。

关键词: 微波炉加热; 提取DNA; 芽孢杆菌; 乳酸菌

Abstract:

The assay was aimed to investigate a fast,simple and efficient method with microwave heating for extracting DNA of Bacillus and Lactobacillus.Treated in a microwave oven at rated power (800 W) for a certain time (40 to 150 s),then centrifuged for supernatant.The supernatant was transferred to a sterile tube for PCR testing and agarose gel electrophoresis.Sequencing was carried out after determining the purity and yield.The results showed that heating for 40 to 150 s at 800 W microwave power conditions could effectively extract DNA of Bacillus and Lactobacillus.The obtained DNA quality was good (D260 nm/D280 nm was 1.8 to 2.1).The DNA concentration in line with the PCR testing requirements;Sequencing results was satisfied with the routine molecular biological research.The microwave extraction method had simple,fast and efficient features,and a wide range of practical,which provided a simple means for rapid molecular assay.

Key words: microwave heating; DNA extraction; Bacillus; Lactobacillus

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