COMET Assay

Please let me know more about Single Cell Gel Electrophoresis / Comet Assay For Rapid Genotoxicity Assessment protocols prepared by Industrial Toxicology Research Centre?

[Kiran Chandar]

I will let you know about Single Cell Gel Electrophoresis / Comet Assay For Rapid Genotoxicity Assessment protocols prepared by Industrial Toxicology Research Centre so that you can check it easily.

The single cell gel electrophoresis (SCGE)/ comet assay, developed by N.P. Singh1, combines the simplicity of biochemical techniques for detecting DNA single strand breaks.

The advantages of the SCGE technique include:
(1) The collection of data at the level of the individual cell, allowing for more robust types of statistical analyses

(2) The need for small numbers of cells per sample (<10,000)

(3) Its sensitivity for detecting DNA damage

(4) That virtually any eukaryotic cell population is amenable to analysis.

I. Preparation of Reagents
Materials: Supplier (Catalogue Number):

Dimethylsulfoxide (DMSO) - Qualigens (CPW59)

Disodium EDTA - HiMedia (RM1370)

Ethidium Bromide - Sigma (E-8751)

Histopaque - Sigma (1077-1)

Phosphate Buffered Saline (PBS) (Ca++, Mg++ free) - HiMedia (TS1006)

Sodium Chloride (NaCl) - Ranbaxy Rankem (S0160)

Sodium Hydroxide (NaOH) - BDH-Merck (89021)

Triton X-100 - HiMedia (RM 845)

Trizma Base - Spectrochem (042061)


Single Cell Gel Electrophoresis / Comet Assay For Rapid Genotoxicity Assessment protocols







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Address:-
Developmental Toxicology Division
Industrial Toxicology Research Centre
P.O. Box – 80, M.G. Marg, Lucknow – 226 001
U.P., India

Phone:-
(91) 522-2213618; Fax: (91) 522-2228227, 2211547