Quantitation of single- and double-strand DNA breaks in vitro and in vivo

Ron Kohen; Moshe Szyf; Mordechai Chevion

doi:10.1016/0003-2697(86)90019-9

Quantitation of single- and double-strand DNA breaks in vitro and in vivo

Ron Kohen^*, Moshe Szyf, Mordechai Chevion

^*Corresponding author for this work

School of Pharmacy

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Abstract

This communication describes a rapid and convenient procedure for quantitation of strand breaks in bacterial DNA, both in vitro and in vivo, using agarose gel electrophoresis. The electrophoretic determination of single strand breaks is carried out in alkaline medium, followed by renaturation of the gel and intercalation of the fluorescent dye, ethidium bromide. Double-strand breaks are determined by electrophoresis in neutral medium containing the dye. The distribution of DNA fragment sizes, the determination of the number-average molecular weight, the quantitation of the average number of DNA breaks per molecule, and the ratio between the single- and double-strand breaks are evaluated from microdensitometric scanning of the gels. The application of this analysis to damage caused by a combination of ascorbate and copper is demonstrated.

Original language	English
Pages (from-to)	485-491
Number of pages	7
Journal	Analytical Biochemistry
Volume	154
Issue number	2
DOIs	https://doi.org/10.1016/0003-2697(86)90019-9
State	Published - 1 May 1986

Bibliographical note

Funding Information:
This work has been supported by a grant from Jnited States-Israel Binationai Science Foundation.

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10.1016/0003-2697(86)90019-9

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title = "Quantitation of single- and double-strand DNA breaks in vitro and in vivo",

abstract = "This communication describes a rapid and convenient procedure for quantitation of strand breaks in bacterial DNA, both in vitro and in vivo, using agarose gel electrophoresis. The electrophoretic determination of single strand breaks is carried out in alkaline medium, followed by renaturation of the gel and intercalation of the fluorescent dye, ethidium bromide. Double-strand breaks are determined by electrophoresis in neutral medium containing the dye. The distribution of DNA fragment sizes, the determination of the number-average molecular weight, the quantitation of the average number of DNA breaks per molecule, and the ratio between the single- and double-strand breaks are evaluated from microdensitometric scanning of the gels. The application of this analysis to damage caused by a combination of ascorbate and copper is demonstrated.",

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AB - This communication describes a rapid and convenient procedure for quantitation of strand breaks in bacterial DNA, both in vitro and in vivo, using agarose gel electrophoresis. The electrophoretic determination of single strand breaks is carried out in alkaline medium, followed by renaturation of the gel and intercalation of the fluorescent dye, ethidium bromide. Double-strand breaks are determined by electrophoresis in neutral medium containing the dye. The distribution of DNA fragment sizes, the determination of the number-average molecular weight, the quantitation of the average number of DNA breaks per molecule, and the ratio between the single- and double-strand breaks are evaluated from microdensitometric scanning of the gels. The application of this analysis to damage caused by a combination of ascorbate and copper is demonstrated.

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Quantitation of single- and double-strand DNA breaks in vitro and in vivo

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