PMID: 4881972Nov 1, 1968Paper

Characterization of rapidly labelled ribonucleic acid in Escherichia coli by deoxyribonucleic acid-ribonucleic acid hybridization

The Biochemical Journal
G H Pigott, J E Midgley

Abstract

1. Rapidly labelled RNA from Escherichia coli K 12 was characterized by hybridization to denatured E. coli DNA on cellulose nitrate membrane filters. The experiments were designed to show that, if sufficient denatured DNA is offered in a single challenge, practically all the rapidly labelled RNA will hybridize. With the technique employed, 75-80% hybridization efficiency could be obtained as a maximum. Even if an excess of DNA sites were offered, this value could not be improved upon in any single challenge of rapidly labelled RNA with denatured E. coli DNA. 2. It was confirmed that the hybridization technique can separate the rapidly labelled RNA into two fractions. One of these (30% of the total) was efficiently hybridized with the low DNA/RNA ratio (10:1, w/w) used in tests. The other fraction (70% of the total) was hybridized to DNA at low efficiencies with the DNA/RNA ratio 10:1, and was hybridized progressively more effectively as the amount of denatured DNA was increased. A practical maximum of 80% hybridization of all the rapidly labelled RNA was first achieved at a DNA/RNA ratio 210:1 (+/-10:1). This fraction was fully representative of the rapidly labelled RNA with regard to kind and relative amount of materials hybri...Continue Reading

Citations

Jul 19, 1969·Nature·P G Jenkins, W T Drabble
Jan 1, 1972·Humangenetik·K Bross, W Krone
Dec 30, 1972·Journal of Molecular Biology·P Donini
Mar 14, 1970·Journal of Molecular Biology·M Adesnik, C Levinthal

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