Northern hybridization analysis of RNA using diethylpyrocarbonate-treated nonfat milk.
A simple and relatively inexpensive method for hybridizing RNA that is immobilized on nitrocellulose to a radioactive DNA probe is presented. The procedure, a modification of the Bovine Lacto Transfer Technique Optimizer method of Johnson et al. (1984) Gene Anal. Tech. 1, 3-8, uses nonfat milk which has been treated with the RNase inhibitor, diethylpyrocarbonate (DEPC), to saturate nonspecific binding sites on nitrocellulose paper, and to wash off unbound radioactivity. The DEPC-nonfat milk hybridization technique is faster, less costly, and more reliable than standard Northern blot hybridization conditions, yielding sharp, clear bands of RNA on autoradiographs with virtually no background reactivity.
D000818:Animals; D001345:Autoradiography; D002417:Cattle; D003101:Collodion; D004047:Diethyl Pyrocarbonate; D008297:Male; D008892:Milk; D009693:Nucleic Acid Hybridization; D012313:RNA / Q000032:analysis*; D051381:Rats; D011919:Rats, Inbred Strains; D012260:Ribonucleases / Q000037:antagonists & inhibitors
D012313:RNA; D003101:Collodion; D012260:Ribonucleases; D004047:Diethyl Pyrocarbonate