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Comet Assay to detect DNA damage (CAT#: STEM-CBT-0011-WXH)

Introduction

The comet assay (single-cell gel electrophoresis) is a simple method for measuring deoxyribonucleic acid (DNA) strand breaks in eukaryotic cells. Cells embedded in agarose on a microscope slide are lysed with detergent and high salt to form nucleoids containing supercoiled loops of DNA linked to the nuclear matrix. Electrophoresis at high pH results in structures resembling comets, observed by fluorescence microscopy; the intensity of the comet tail relative to the head reflects the number of DNA breaks.
At present, the common comet assay detection methods are divided into the following two types: under alkaline treatment conditions, DNA denatures, and all DNA damage including single-strand and double-strand breaks can be detected; under neutral treatment conditions, DNA maintains double-strand conformation, Only double-strand breaks can be detected.




Applications

• Rapid detection of single-cell DNA damage.
• Screening of carcinogenic and mutagenic genotoxic substances, monitoring of genotoxic substances in environmental media.
• Study the mechanism of carcinogenic action of compounds.
• Determining the degree of DNA fragmentation in sperm cells.

Procedure

1. Cell Culture and Treatment
2. Cell Collection
3. Film production and glue laying
4. Cell Lysis
5. DNA unwinding
6. Single Cell Electrophoresis
7. Rinsing and dyeing
8. Fluorescence microscopy results observation

Notes

Customer provided:
1. Cell type and name
2. Detection time node

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