Prima V I, Platonov O M, Gazarian K G
Biokhimiia. 1980 Mar;45(3):498-506.
A method for direct registration of the first derivative of DNA reassociation kinetics in spectrophotometric cuvettes is proposed. Using this method, the rat liver DNA obtained by chromatography on hydroxyapatite was studied. The differential curves of reassociation and changes in hyperchromic spectra revealed the presence in the rat genome of 4 types of repeating sequences differing in their GC-content. A scheme of preparative separation of DNA with respect to the reassociation rate was developed. Three types of sequences repeating 3.10(5), 600 and 1 times in the genomes with the GC-content of 50, 42, and 33%, respectively, were obtained in a homogeneous state. The advantages of the differential method of registration of the kinetic curves for the study of DNA structure and calculation of fractionation schemes are discussed.
提出了一种在分光光度比色皿中直接记录DNA重缔合动力学一阶导数的方法。利用该方法,对通过羟基磷灰石层析获得的大鼠肝脏DNA进行了研究。重缔合的微分曲线和增色光谱的变化揭示了大鼠基因组中存在4种GC含量不同的重复序列。制定了根据重缔合速率对DNA进行制备性分离的方案。分别获得了基因组中重复3.10(5)、600和1次且GC含量分别为50%、42%和33%的三种类型序列的均一状态。讨论了用于研究DNA结构和计算分级分离方案的动力学曲线微分记录法的优点。