Hybridization of polyuridylic acid to human DNA immobilized onto nitrocellulose filters

The level of deoxyadenylate (dA) regions in human DNA was estimated from formation of poly(U)-poly(dA) triplexes on nitrocellulose filters that were RNAase resistant. The (dA) rich sequences were determined following mild ribonuclease treatment of the poly(U)-DNA hybrids (5 μg/ml at 25°C for 30 min)...

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Bibliographic Details
Published in:Nucleic acids research 1975-07, Vol.2 (7), p.1163-1176
Main Authors: Pero, Ronald W., Bryngelsson, Tomas, Bryngelsson, Carl, Deutsch, Adam, Nordén, Åke, Norgren, Allan
Format: Article
Language:English
Subjects:
Adenine Nucleotides - analysis
Breast - analysis
Centrifugation, Density Gradient
Chromatography
DNA - analysis
DNA - blood
DNA - isolation & purification
Female
Humans
Hydroxyapatites
Lymphocytes - analysis
Nucleic Acid Hybridization
Pancreas - enzymology
Placenta - analysis
Poly U
Pregnancy
Ribonucleases
Ultrafiltration
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Summary:The level of deoxyadenylate (dA) regions in human DNA was estimated from formation of poly(U)-poly(dA) triplexes on nitrocellulose filters that were RNAase resistant. The (dA) rich sequences were determined following mild ribonuclease treatment of the poly(U)-DNA hybrids (5 μg/ml at 25°C for 30 min), where as exhaustive ribonuclease treatment (5 μg/ml at 25°C for 6 hr) estimated the more (dA) pure sequences. The level of (dA) rich regions was 0.39% of the DNA and for the more (dA) pure regions it was 0.07%. The (dA) regions were widely distributed throughout human DNA regardless of base composition or sequence repetition. However, a concentration of (dA) regions into main band CsCl gradient fractions of DNA and into repeated DNA was observed.
ISSN:0305-1048
1362-4962
DOI:10.1093/nar/2.7.1163