Rapid isolation of carcinogen-bound DNA and RNA by hydroxyapatite chromatography.
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.
D000818:Animals; D001665:Binding Sites; D002273:Carcinogens / Q000302:isolation & purification* / Q000378:metabolism; D002417:Cattle; D002845:Chromatography / Q000379:methods; D004247:DNA / Q000302:isolation & purification* / Q000378:metabolism; D006868:Hydrolysis; D006882:Hydroxyapatites; D008099:Liver / Q000032:analysis; D008297:Male; D012313:RNA / Q000302:isolation & purification* / Q000378:metabolism; D051381:Rats; D013950:Thymus Gland / Q000032:analysis
D002273:Carcinogens; D006882:Hydroxyapatites; D012313:RNA; D004247:DNA