Assay of DNA-RNA hybrids by S1 nuclease digestion and adsorption to DEAE-cellulose filters.
A fast and accurate assay procedure for DNA-RNA hybrids is described in which exhaustive digestion of unhybridized DNA with S1 nuclease is followed by binding of hybrids to filter discs of DEAE-cellulose. The digested DNA can be efficiently washed from the filters so that background levels of 0.1-0.2% of input tracer DNA can be achieved, in contrast to the much higher (approximately 1-5%) backgrounds obtained using TCA precipitation procedures. Short duplexes, as small as 36 nucleotides in length, which are inefficiently bound to hydroxyapatite, are quantitatively bound to the DEAE-cellulose filters.
D000818:Animals; D001921:Brain; D002482:Cellulose; D004247:DNA; D003851:Deoxyribonucleases; D007700:Kinetics; D008722:Methods; D051379:Mice; D009693:Nucleic Acid Hybridization; D012313:RNA; D012260:Ribonucleases
D012313:RNA; D002482:Cellulose; D004247:DNA; D003851:Deoxyribonucleases; D012260:Ribonucleases