Assay of DNA-RNA hybrids by S1nuclease digestion and adsorption to DEAE-cellulose filters
- 1 January 1978
- journal article
- Published by Oxford University Press (OUP) in Nucleic Acids Research
- Vol. 5 (6), 2033-2038
- https://doi.org/10.1093/nar/5.6.2033
Abstract
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.Keywords
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