SYBR Green I staining of pulsed field agarose gels is a sensitive and inexpensive way of quantitating DNA double-strand breaks in mammalian cells

Anne E. Kiltie; Anderson J. Ryan

doi:10.1093/nar/25.14.2945

SYBR Green I staining of pulsed field agarose gels is a sensitive and inexpensive way of quantitating DNA double-strand breaks in mammalian cells

Anne E. Kiltie, Anderson J. Ryan^*

^*Corresponding author for this work

The Rowett Institute of Nutrition and Health

University of Manchester

Research output: Contribution to journal › Article › peer-review

44 Citations (Scopus)

Abstract

Pulsed field gel electrophoresis (PFGE) is widely used to measure DNA double strand breaks (dsb). The DNA of cultured cells can be prelabelled with radioactivity, which helps greatly in detection and quantitation of DNA dsb. However, this approach cannot be used with non-cycling cells from biopsy material. We describe a method which uses SYBR Green I to stain DNA in dried agarose gels. DNA is detected and analysed using readily available camera equipment and image analysis software. This method is as sensitive as [³H]thymidine prelabelling of cells and allows DNA dsb to be measured simply and economically in non-cycling cells.

Original language	English
Pages (from-to)	2945-2946
Number of pages	2
Journal	Nucleic Acids Research
Volume	25
Issue number	14
DOIs	https://doi.org/10.1093/nar/25.14.2945
Publication status	Published - 15 Jul 1997

Bibliographical note

Funding Information:
A.E.K. is supported on a grant (Principle Investigator J.H.Hendry) from the Christie Hospital NHS Trust Endowment Fund. We wish to thank Professor Jolyon Hendry, Dr Geoff Margison and Mr Tim Ward for their help throughout this work.

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abstract = "Pulsed field gel electrophoresis (PFGE) is widely used to measure DNA double strand breaks (dsb). The DNA of cultured cells can be prelabelled with radioactivity, which helps greatly in detection and quantitation of DNA dsb. However, this approach cannot be used with non-cycling cells from biopsy material. We describe a method which uses SYBR Green I to stain DNA in dried agarose gels. DNA is detected and analysed using readily available camera equipment and image analysis software. This method is as sensitive as [3H]thymidine prelabelling of cells and allows DNA dsb to be measured simply and economically in non-cycling cells.",

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SYBR Green I staining of pulsed field agarose gels is a sensitive and inexpensive way of quantitating DNA double-strand breaks in mammalian cells

Abstract

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