生鲜牛乳总微生物基因组DNA的提取.PDFVIP

3 5 Vol. 3 No. 5 2012 10 Journal of Food Safety and Quality Oct. , 2012 DNA 1 2* 3 2 1* 龙丹丹 , 张敏爱 ,王子亮 , 张建军 , 杨秀清 (1. , 030006; 2. 030024; 3. , 100062) 摘 要: 目的 DNA 方法 , SDS- K , , DNA 结果 NET(Tris•HCl, EDTA, NaCl), K DNA , ; NCT(NaCl, CaCl2, Tris•HCl), PCR RNA (KAc), , , DNA SDS-K DNA 关键词: ; DNA; PCR Extraction of total microbial genomic DNA in raw milk 1 2* 3 2 1* LONG Dan-Dan , ZHANG Min-Ai , WANG Zi-Liang , ZHANG Jian-Jun , YANG Xiu-Qing (1. Institute of Biotechnology , Shanxi University, Taiyuan 030006, China; 2. Shanxi Entry-Exit Inspection and Quarantine Bureau of PRC, Taiyuan 030024, China; 3. National Institute for Public Health and the Environment, Beijing 100062, China) ABSTRACT: Objective To establish an efficient and rapid method for the genomic DNA extraction of mi- crobe from raw milk. Methods Genomic DNA of microbe was prepared after the cell lysis by SDS-proteinase K method and removing protein by phenol-chloroform and potassium acetate. Results Genomic DNA of mi- crobe, which was obtained by using of proteinase K and NET(Tris•HCl, EDTA, NaCl) as buffer, showed some advantages of high purity and yield and time saving. Comparatively, the specificity and yield of polymerase chain reaction(PCR) products were lower while chosing NCT (NaCl, CaCl2, Tris•HCl) as buffer. RNAse hardly af