使用异丙醇分步沉淀法初步纯化质粒 DNA。
Primary Purification of Plasmid DNA Using Differential Isopropanol Precipitation.
机构信息
Department of Bioengineering, Institute for Bioengineering and Biosciences, Instituto Superior Técnico, Universidade de Lisboa, Lisbon, Portugal.
Institut National des Sciences Appliquées de Rouen, Saint-Étienne-du-Rouvray, France.
出版信息
Methods Mol Biol. 2021;2197:151-165. doi: 10.1007/978-1-0716-0872-2_8.
A method for the intermediate recovery of plasmid DNA (pDNA) from alkaline lysates is described that comprises differential isopropanol precipitation steps. In a first low-cut precipitation, a smaller amount of isopropanol (20% v/v) is used so that only high molecular weight RNA precipitates. After solid liquid separation, a high-cut precipitation is performed by bringing isopropanol concentration to 70% v/v to precipitate pDNA. Tests made with lysates show that the differential precipitation increases purity threefold compared to the conventional one-step precipitation at 70% v/v without affecting pDNA recovery (>80%).
介绍了一种从中性裂解物中回收质粒 DNA（pDNA）的方法，该方法包括分步异丙醇沉淀。在第一次低浓度沉淀中，使用较少量的异丙醇（20%v/v），以便仅沉淀高分子量 RNA。固体液体分离后，将异丙醇浓度提高到 70%v/v 进行高浓度沉淀，以沉淀 pDNA。用裂解物进行的测试表明，与传统的 70%v/v 一步沉淀相比，分步沉淀可将纯度提高三倍，而不影响 pDNA 的回收（>80%）。
Zotero 插件