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PMID: 673844 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Assay of DNA-RNA hybrids by S1 nuclease digestion and adsorption to DEAE-cellulose filters.

Nucleic acids research ·Vol. 5 ·No. 6 ·1978-06-00 ·Pages 2033-8

Maxwell IH, Van Ness J, Hahn WE

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.

MeSH Terms
Animals Brain Cellulose DNA Deoxyribonucleases Kinetics Methods Mice Nucleic Acid Hybridization RNA Ribonucleases
Chemicals
RNA Cellulose DNA Deoxyribonucleases Ribonucleases
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Maxwell I H
Van Ness J
Hahn W E
References (11)
11 references, click to expand
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/5.6.2033
Published
1978-06-00
Pages
2033-8
Language
English
Region
England
NLM ID
0411011
PMCID
PMC342142
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/5.6.2033 PMC Full Text View journal details
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