Beland F A, Dooley K L, Casciano D A
J Chromatogr. 1979 Jun 1;174(1):177-86. doi: 10.1016/s0021-9673(00)87048-x.
Carcinogen-bound DNA and RNA are conveniently isolated by solvent extraction and hydroxyapatite (HAP) chromatography. Tissue is suspended in 8 M urea-0.24 M sodium phosphate-1% sodium dodecyl sulfate-10 mM EDTA, pH 6.8 (MUP-SDS-EDTA) and extracted with chloroform-isoamyl alcohol-phenol (24:1:25; CIP) to remove protein. RNA and DNA are separated by passing the aqueous solution through an HAP column; RNA is eluted with MUP, DNA with 0.48 M sodium phosphate, pH 6.8. Examples presented are: (1) calf thymus DNA that has been reacted with N-acetoxy-2-acetylaminofluorene (N-OAc-AAF), (2) isolated intact rat hepatocytes incubated with N-hydroxy-AAF and (3) livers from Sprague-Dawley rats treated with N-hydroxy-AAF.
与致癌物结合的DNA和RNA可通过溶剂萃取和羟基磷灰石（HAP）色谱法方便地分离出来。将组织悬浮于pH 6.8的8M尿素-0.24M磷酸钠-1%十二烷基硫酸钠-10mM乙二胺四乙酸（MUP-SDS-EDTA）中，并用氯仿-异戊醇-苯酚（24:1:25；CIP）萃取以去除蛋白质。通过使水溶液通过HAP柱来分离RNA和DNA；RNA用MUP洗脱，DNA用pH 6.8的0.48M磷酸钠洗脱。给出的例子有：（1）与N-乙酰氧基-2-乙酰氨基芴（N-OAc-AAF）反应的小牛胸腺DNA，（2）用N-羟基-AAF孵育的分离完整大鼠肝细胞，以及（3）用N-羟基-AAF处理的Sprague-Dawley大鼠的肝脏。
