Catalog #

Product Name

Size

4250-050-K

CometAssay? Kit (25 X 2 well slides)

50 samples

APPLICATIONS:                                                                       

• Applicable for the analysis of either single strand or double strand DNA breaks                                                                                
• Study cellular responses to DNA damage
• Screen for inhibitors of DNA repair
• Test chemicals for toxicity
• Screen for environmental mutagens
• Probe for specific types of DNA damage using DNA repair glycosylases

Trevigen's CometAssay?, or single cell gel electrophoresis assay, provides a simple and effective method for evaluating DNA damage in cells. The principle of the assay is based upon the ability of denatured, cleaved DNA fragments to migrate out of the nucleoid under the influence of an electric field, whereas undamaged DNA migrates slower and remains within the confines of the nucleoid when a current is applied. Evaluation of the DNA "comet" tail shape and migration pattern allows for assessment of DNA damage. The Neutral CometAssay? is typically used to detect double-stranded breaks, whereas the Alkaline CometAssay? is more sensitive, and is used to detect smaller amounts of damage includingsingle and double-stranded breaks.

ASSAY DESIGN FOR ALKALINE COMET:

Step 1: Cells mixed with low melting agarose are immobilized on specially treated slides and lysed.

• Membranes and histones removed from the DNA of individually isolated cells.

Step 2: Samples are treated with alkali to unwind and denature the DNA.

• For detection of single and double-stranded breaks.

Step 3: Following alkaline electrophoresis slides are stained and visualized by fluorescent microscopy.

• Increase in DNA damage results in smaller fragments migrating faster in an electric field.
• Quantitative measurements of DNA damage