Nedeva Trayana, Petrova Ventzislava, Hristozova Tsonka, Kujumdzieva Anna
Department of General and Industrial Microbiology, Faculty of Biology, The Sofia University St. Kliment Ohridski, Bulgaria.
Z Naturforsch C J Biosci. 2002 Sep-Oct;57(9-10):960-1. doi: 10.1515/znc-2002-9-1036.
A modified, rapid and inexpensive method for preparation of mitochondrial DNA (mtDNA), suitable for molecular analysis is proposed. It comprises batch cultivation of Saccharomyces cerevisiae strain NBIMCC 583 on a simple nutrient medium at 28 degrees C; permeabialization of cells from late exponential growth phase with cetyltrimethylamonnium bromide, mechanical disintegration of the cell wall; preparation of a mitochondrial fraction and subsequent isolation and purification of mtDNA. The amount and the purity of the obtained mtDNA have been checked and its application for molecular analysis proven. The main advantages of the proposed procedure for isolation of mtDNA are introduction of simple nutrient medium, replacement of the enzymatic lysis of the cell wall by the cheaper mechanical one, avoidance of ultracentrifugation steps and use of harmful chemical substances.
本文提出了一种改良的、快速且廉价的制备线粒体DNA（mtDNA）的方法，适用于分子分析。该方法包括在28℃的简单营养培养基上分批培养酿酒酵母菌株NBIMCC 583；用十六烷基三甲基溴化铵使处于指数生长后期的细胞透化，机械破碎细胞壁；制备线粒体组分，随后分离和纯化mtDNA。已对所得mtDNA的量和纯度进行了检测，并证明了其在分子分析中的应用。所提出的mtDNA分离方法的主要优点包括采用简单营养培养基、用更廉价的机械方法替代细胞壁的酶解、避免超速离心步骤以及使用有害化学物质。
