To assess the type of DNA damage induced by a putative mutagen, drug, or treatment regimen, cells are harvested after treatment and immobilized in a layer of low melting point agarose on the FLARE Slide.
The cells are gently lysed and subsequently incubated with a kit-specific enzyme which recognizes a particular type of DNA damage and cleaves the DNA. After treatment with alkali to denature the DNA, the samples are submitted to neutral gel electrophoresis, and stained using SYBR(r) Green I. Denatured, cleaved DNA fragments migrate out of the cell during electrophoresis, while undamaged, supercoiled DNA remains within the confines of the nuclear cell membrane.
The extent of the DNA damage can be assessed qualitatively, or quantitatively using image analysis software to calculate tail length and tail moment.
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