Kiper M
Institut für Botanik der TU, Hannover, German Federal Republic.
Anal Biochem. 1978 Nov;91(1):70-4. doi: 10.1016/0003-2697(78)90816-3.
During the renaturation of DNAs large networks build up which cannot be eluted from hydroxyapatite under standard fractionation conditions (60 degrees C, 0.5 M PB). This is a serious problem especially in plant-DNA renaturation studies as hyperpolymers may comprise more than half of the renatured DNA mass even at moderately long initial fragment lengths and low C0t values. Utilizing the acid solubility of hydroxyapatite a method is outlined which will recover the total double-stranded DNA fraction and will prepare the column for the next fractionation in one quick operation. As the method is time saving compared to the standard hydroxyapatite fractionation procedure its general application may prove to be useful.
在DNA复性过程中会形成大的网络结构,在标准分级分离条件下(60℃,0.5M磷酸缓冲液)无法从羟基磷灰石上洗脱下来。这是一个严重的问题,尤其是在植物DNA复性研究中,因为即使在初始片段长度适中且C0t值较低的情况下,超聚合物可能占复性DNA总量的一半以上。利用羟基磷灰石的酸溶性,本文概述了一种方法,该方法可以在一次快速操作中回收双链DNA的总级分,并为下一次分级分离准备柱子。由于该方法与标准的羟基磷灰石分级分离程序相比节省时间,其广泛应用可能会很有用。