Rapid isolation of carcinogen-bound DNA and RNA by hydroxyapatite chromatography
Beland, F.A.; Dooley, K.L.; Casciano, D.A.
Journal of Chromatography 174(1): 177-186
1979
ISSN/ISBN: 0021-9673 PMID: 94918 Document Number: 143903
Carcinogen-bound DNA and RNA were conveniently isolated by solvent extraction and hydroxyapatite (HAP) chromatography. Tissue was 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-isoamylalcohol-phenol (24:1:25; CIP) to remove protein. RNA and DNA were 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 were: calf thymus DNA that was reacted with N-acetoxy-2-acetylaminofluorene (N-OAc-AAF), isolated intact rat hepatocytes incubated with N-hydroxy-AAF and livers from Sprague-Dawley rats treated with N-hydroxy-AAF.