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Propidium lodide and S1 nuclease: Tools for studying DNA reassociation kinetics

✍ Scribed by William Davies; Øivind Nordby

Publisher: Elsevier Science
Year: 1985
Tongue: English
Weight: 449 KB
Volume: 146
Category: Article
ISSN: 0003-2697
DOI: 10.1016/0003-2697(85)90562-7

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✦ Synopsis

A method for the determination of the amount of double-stranded DNA in a reassociation mixture is described. Reassociated DNA resistant to Sl nuclease digestion is measured fluorometrically using propidium iodide. A direct comparison is made between this method and an established method in which radiolabeled Escherichia coli DNA resistant to Sl digestion is measured by scintillation counting after separation of nucleotides by Sephadex G-100 chromatography. Reassociation curves determined for calf thymus and E. coli DNA are presented.

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✍ Chanchal Sadhu; Santanu Datta; K.P. Gopinathan 📂 Article 📅 1984 🏛 Elsevier Science 🌐 English ⚖ 403 KB

The optimal parameters in the use of nuclease S1 in DNA reassociation kinetics in the presence of formamide have been determined. The conditions are especially suitable for the study of DNA rich in mole percent GC. A 10-fold dilution of the reassociation samples leading to a decrease in both NaCl an