一种简单的获得高纯度沃尔巴克氏体共生菌 DNA 用于基因组测序的方法。
A simple protocol to obtain highly pure Wolbachia endosymbiont DNA for genome sequencing.
机构信息
School of Biological Sciences, The University of Queensland, Brisbane, Qld 4072, Australia.
出版信息
J Microbiol Methods. 2011 Jan;84(1):134-6. doi: 10.1016/j.mimet.2010.10.019. Epub 2010 Oct 31.
Most genome sequencing projects using intracellular bacteria face difficulties in obtaining sufficient bacterial DNA free of host contamination. We have developed a simple and rapid protocol to isolate endosymbiont DNA virtually free from fly and mosquito host DNA. We purified DNA from six Wolbachia strains in preparation for genome sequencing using this method, and achieved up to 97% pure Wolbachia sequence, even after using frozen insects. This is a significant improvement for future Wolbachia and other endosymbiont genome projects.
大多数使用胞内细菌的基因组测序项目都面临着难以获得无宿主污染的足够细菌 DNA 的困难。我们开发了一种简单而快速的方法来分离实质上不含果蝇和蚊子宿主 DNA 的共生体 DNA。我们使用该方法从六种沃尔巴克氏体菌株中纯化了 DNA，为基因组测序做准备，即使使用冷冻昆虫也能获得高达 97%的纯沃尔巴克氏体序列。这对于未来的沃尔巴克氏体和其他共生体基因组项目是一个重大改进。
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