Naktinis V I, Maleeva N E, San'ko D F, Mirzabekov A D
Biokhimiia. 1977 Oct;42(10):1783-90.
Two general methods for the isolation of DNA from various sources based on the use of cetyltrimethylammonium bromide (cetavlon, CTA-Br) are described. Cetavlon is a strong cationic detergent precipitating DNA from diluted salt solutions. Cells are lysed and cellular components are dissolved in the presence of cetavlon, 5 M urea, 0.1 M EDTA and 2 M NaCl (KCl). In the first method pure DNA is precipitated in the form of CTA-salt by direct dilution of the lysate to bring the concentration of NaCl (KCl) down to 0.5 M after the removal of the main part of proteins by deproteinization with chloroform. In the first method pure DNA is precipitated in the form of CTA-salt by direct dilution of the lysate to bring the concentration of NaCl (KCl) down to 0.5 M after the removal of the main part of proteins by deproteinization with chloroform. In the second method DNA is purified on the hydroxyapatite column after cell lysis and the removal of cell debris by centrifugation. Both methods are suitable for rapid isolation of pure DNA from various sources with recovery about 80% and average molecular weight 20-10(6) and higher without use of ribonuclease, pronase and amylase.
本文描述了两种基于使用十六烷基三甲基溴化铵（西曲溴铵，CTA-Br）从各种来源分离DNA的通用方法。西曲溴铵是一种强阳离子去污剂，可从稀释的盐溶液中沉淀DNA。细胞在西曲溴铵、5M尿素、0.1M乙二胺四乙酸（EDTA）和2M氯化钠（氯化钾）存在的情况下被裂解，细胞成分被溶解。在第一种方法中，通过用氯仿脱蛋白去除大部分蛋白质后，直接稀释裂解液使氯化钠（氯化钾）浓度降至0.5M，从而以CTA盐的形式沉淀出纯DNA。在第二种方法中，细胞裂解后通过离心去除细胞碎片，然后在羟基磷灰石柱上纯化DNA。两种方法都适用于从各种来源快速分离纯DNA，回收率约为80%，平均分子量为20 - 10(6)及更高，且无需使用核糖核酸酶、链霉蛋白酶和淀粉酶。
