Evaluation Of COMET SCGE Assay Kit

City of Hope Cancer Research Hospital
Department of Cancer Genetics and Epigenetics
Assistant Research Professor

Overall

Quality of Results

Ease-of-Optimization

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Company:

Enzo

Product Name:

COMET SCGE Assay Kit

Catalog Number:

ADI-900-166

The general aim of the research is to study the function of a gene in DNA damage repairs in cancer cells. The specific goals of the experiment were to examine a role of a gene in DNA damage repair in MDA-MB-231 triple negative breast cancer cell lines. Cells after scramble (left) or gene-specific (right) shRNA knock-down were subjected to Enzo Comet assay. This kit makes the complicated comet assay easy and handy to handle. ­

Experimental Design and Results Summary

Application

Single Cell Electrophoresis Assay

Starting Material

MDA-MB-231 triple negative breast cancer cell lines

Protocol Overview

Cells were collected and rinsed twice with ice cold PBS and combined with LMAgarose at 37˚C at a ratio of 1: 10 (v/v) and immediately pipetted onto slides. For cell lysis, the slides were immersed in in prechilled Lysis Solution at 4˚C, for 30 to 60 minutes, and then in freshly prepared Alkaline Solution, pH>13, for 20 to 60 minutes at room temperature, in the dark. The slides were subjected to electrophoresis at 15 V for 25 minutes (0.6 V/cm), and stained in CYGREEN® Nucleic Acid Dye for 30 minutes. Images were taken with a fluorescence microscope and analyzed with the CometScore software.

Tips

Resuspend cells at 2 x 105 cells/ml but not 1 x 105 cells/ml in ice cold 1X PBS (Ca2+ and Mg2+ free). It will give the signals from more cells when taking images.

Results Summary

The results are excellent. It works all the times.

Additional Notes

None

Image Gallery

Summary

The Good

Easy to use

The Bad

Long procedures

The Bottom Line

Easy, simple and works all the time

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