Lee Y K, Kim H W, Liu C L, Lee H K
Microbiology Laboratory, Korea Ocean Research and Development Institute, Ansan PO Box 29, 425-600, South Korea.
J Microbiol Methods. 2003 Feb;52(2):245-50. doi: 10.1016/s0167-7012(02)00180-x.
We present a simple method for extracting DNA from the marine bacteria Hahella chejuensis, a Streptomyces sp., and a Cytophaga sp. Previously, DNA purification from these strains was hindered by the presence of extracellular materials. In our extraction method, the marine bacteria are lysed by freezing and grinding in liquid nitrogen, and treated with SDS. The extracted DNA is purified using a phenol/chloroform mixture, and precipitated in isopropanol. The extracted DNA is of high quality and suitable for molecular analyses, such as PCR, restriction enzyme digestion, genomic DNA blot hybridization, and genomic DNA library construction. We used this method to extract genomic DNA from several other marine bacteria. Our method is a reproducible, simple, and rapid technique for routine DNA extractions from marine bacteria. Furthermore, the low cost of this method makes it attractive for large-scale studies.
我们提出了一种从海洋细菌济州海杆菌、一种链霉菌属细菌和一种噬纤维菌属细菌中提取DNA的简单方法。此前,这些菌株的DNA纯化因细胞外物质的存在而受阻。在我们的提取方法中,将海洋细菌在液氮中冷冻研磨进行裂解,并用十二烷基硫酸钠(SDS)处理。提取的DNA用苯酚/氯仿混合物纯化,然后在异丙醇中沉淀。提取的DNA质量高,适用于PCR、限制性酶切、基因组DNA印迹杂交和基因组DNA文库构建等分子分析。我们用这种方法从其他几种海洋细菌中提取基因组DNA。我们的方法是一种可重复、简单且快速的从海洋细菌中常规提取DNA的技术。此外,该方法成本低,使其对大规模研究具有吸引力。