The single cell gel electrophoresis (SCGE) or comet assay is a sensitive and rapid technique for quantifying DNA damage and repair from in vivo and in vitro samples of eukaryotic and plant cells. The resulting image that is obtained resembles a “comet” with a distinct head and tail. The head is composed of intact DNA, while the tail consists of single-strand or double-strand DNA breaks. The extent of DNA liberated from the head of the comet is directly proportional to the amount of DNA damage. This is the only technique that directly measures DNA damage in individual cells and thus has rapidly gained importance in the fields of genetic toxicology and human biomonitoring.
Trevigen offers a complete system for the standard comet assay (CometAssay®) providing kits and reagents including our specially treated CometSlides™ and Control Cells. The FLARE™ (Fragment Length Analysis using Repair Enzymes) Assay provides the ability to probe for oxidative DNA damage in conjunction with CometAssay® reagents. In addition to the standard comet assay, Trevigen’s exclusive 96-Well CometChip® System provides a high-throughput platform to simultaneously treat and measure DNA damage on a single slide using an array of non-overlapping cells. Trevigen’s Analysis Software and Electrophoresis System are designed to improve assay performance and work with both CometAssay®, FLARE™, and CometChip® products.
The 96-Well CometChip® System is a high-throughput platform to simultaneously treat and measure DNA damage induced by different treatments, or among different cell types on a single slide using the comet assay. The CometChip® is a consumable consisting of specifically sized micron pores patterned into agarose. 96 separate wells are created by inserting the CometChip® into a reusable magnetically-sealed cassette suitable for tissue culture incubators. Cells added to each well are captured by gravity into micropores and excess cells are aspirated, leaving an array of non-overlapping cells. Multiple experimental conditions are performed in parallel by the addition of different chemicals to respective wells. Once treatment is complete the CometChip® is removed from the cassette and processed using standard alkaline comet conditions and imaging systems. Electrophoresis of the CometChip® is designed to work with Trevigen's CometAssay® Electrophoresis System and reagents.
Comet Analysis Software (1)
The Comet Analysis Software can rapidly evaluate large numbers of cells and generate summary statistics, increasing processivity approximately 10,000% over traditional comet analysis. It is specially engineered to complement and operate as the computational component of Trevigen's unique CometAssay® Electrophoresis System. The power, ease and speed of analysis can be especially appreciated when coupled with Trevigen's exclusive CometChip® High Throughput System. It is compatible with 4X images, an important feature as most commercially available software operates with 10X images only. This factor allows for rapid screening - greater than 300 comets in seconds when using the CometChip®.
Standard Comet Assay (CometAssay®) (20)
In the standard comet assay, cells are immobilized in a bed of low melting point agarose on a specially treated microscope slide. Following gentle cell lysis, and for the alkaline comet assay, samples are treated with alkali to unwind and denature the DNA and hydrolyze sites of damage. The remaining nucleoids are subjected to electrophoresis and subsequent staining with a fluorescent DNA intercalating dye. Trevigen provides a complete standardized system for consistent results which includes CometAssay® Kits, CometSlides™, Alkaline and Neutral CometAssay® Control Cells and a specialized electrophoresis unit.
Contract Services are also available for the Comet Assay.
- CometAssay® Electrophoresis Starter Kits
- CometAssay® Reagent Kits
- CometAssay® Silver Staining
- CometSlides™ and Rack System
- CometAssay® Control Cells
- CometAssay® Reagents
FLARE™ Assay (11)
Trevigen’s FLARE™ (Fragment Length Analysis using Repair Enzymes) Assay Kits provide the ability to assess the type of DNA damage using DNA Repair enzymes when used in combination with the comet assay. Cells harvested after treatment are immobilized in low melting point agarose on a specially treated microscope slide. The cells are gently lysed and then incubated with DNA Repair enzyme to assess the type of damage present. The slides are then immersed in an alkali solution, to unwind and denature the DNA strands, and subjected to electrophoresis and subsequent DNA staining. The type of DNA damage is inferred from the substrate specificity of the DNA Repair enzyme. Trevigen provides a complete standardized system for consistent results which includes Fpg, hOGG1, and Endo III specific FLARE™ Assay Kits containing FLARE™ Slides designed to work with Trevigen’s CometAssay® Electrophoresis System and reagents.