Vingataramin Laurie, Frost Eric H
Département de microbiologie et d'infectiologie, Faculté de médecine et des sciences de la santé, Université de Sherbrooke, Sherbooke, Quebec, Canada.
Biotechniques. 2015 Mar 1;58(3):120-5. doi: 10.2144/000114263. eCollection 2015 Mar.
Guanidine thiocyanate breakage of microorganisms has been the standard initial step in genomic DNA (gDNA) extraction of microbial DNA for two decades, despite the requirement for pretreatments to extract DNA from microorganisms other than Gram-negative bacteria. We report a quick and low-cost gDNA extraction protocol called EtNa that is efficient for bacteria and yeast over a broad range of concentrations. EtNa is based on a hot alkaline ethanol lysis. The solution can be immediately centrifuged to yield a crude gDNA extract suitable for PCR, or it can be directly applied to a silica column for purification.
二十年来，硫氰酸胍裂解微生物一直是从微生物中提取基因组DNA（gDNA）的标准初始步骤，尽管从革兰氏阴性菌以外的微生物中提取DNA需要进行预处理。我们报告了一种快速且低成本的gDNA提取方案，称为EtNa，它在很宽的浓度范围内对细菌和酵母都有效。EtNa基于热碱性乙醇裂解。该溶液可立即离心以产生适用于PCR的粗gDNA提取物，也可直接应用于硅胶柱进行纯化。
