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PMID: 218170 Published · ppublish English Journal Article

Quantitative analysis of specific labelled RNA'S using DNA covalently linked to diazobenzyloxymethyl-paper.

Nucleic acids research ·Vol. 6 ·No. 1 ·1979-01-00 ·Pages 195-203

Stark GR, Williams JG

Abstract

Substantial amounts of DNA (at least 25 microgram per cm2) can be stably bound to diazobenzyloxymethyl (DBM)-paper. Complementary RNA will hybridize to the DNA paper almost completely in 24 hours. Using several different conditions of hybridization and washing, the background of RNA bound non-specifically is very low (between 0.01 and 0.02%) and the efficiency of hybridization is very high (75 to 50% of complementary RNA is bound and retained through the washing procedure). Because the DNA is bound to the paper convalently, it is retained through all the washing and elution steps, and the DNA papers can be re-used many times.

MeSH Terms
Chromatography, Affinity Chromatography, Paper/methods DNA, Viral Nucleic Acid Hybridization RNA/analysis Simian virus 40
Chemicals
DNA, Viral RNA
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Stark G R
Williams J G
References (7)
7 references, click to expand
  1. Interferon inhibits the in vitro accumulation of virus specific RNA in nuclei isolated from SV40 infected cells.
    Biochem Biophys Res Commun. 1977 Mar 7;75(1):172-8 PMID: 192228
  2. Method for detection of specific RNAs in agarose gels by transfer to diazobenzyloxymethyl-paper and hybridization with DNA probes.
    Proc Natl Acad Sci U S A. 1977 Dec;74(12):5350-4 PMID: 414220
  3. Transcription of Simian virus 40. I. Separation of the strands of SV40 DNA and hybridization of the separated strands to RNA extracted from lytically infected and transformed cells.
    J Mol Biol. 1972 Sep 14;70(1):57-71 PMID: 4342195
  4. Loss of simian virus 40 DNA-RNA hybrids from nitrocellulose membranes; implications for the study of virus--host DNA interactions.
    Proc Natl Acad Sci U S A. 1972 Aug;69(8):2160-4 PMID: 4340749
  5. Selective extraction of polyoma DNA from infected mouse cell cultures.
    J Mol Biol. 1967 Jun 14;26(2):365-9 PMID: 4291934
  6. Transcription of ribonucleic acid in isolated mouse myeloma nuclei.
    Biochemistry. 1973 Aug 28;12(18):3440-6 PMID: 4731189
  7. A quantitative assay for DNA-RNA hybrids with DNA immobilized on a membrane.
    J Mol Biol. 1965 Jul;12(3):829-42 PMID: 4955314
Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
Published
1979-01-00
Pages
195-203
Language
English
Region
England
NLM ID
0411011
PMCID
PMC327682
Subset
IM
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