High-throughput comet assay using 96 minigels.

Abstract:

:The single-cell gel electrophoresis--the comet assay--has proved to be a sensitive and relatively simple method that is much used in research for the analysis of specific types of DNA damage, and its use in genotoxicity testing is increasing. The efficiency of the comet assay, in terms of number of samples processed per experiment, has been rather poor, and both research and toxicological testing should profit from an increased throughput. We have designed and validated a format involving 96 agarose minigels supported by a hydrophilic polyester film. Using simple technology, hundreds of samples may be processed in one experiment by one person, with less time needed for processing, less use of chemicals and requiring fewer cells per sample. Controlled electrophoresis, including circulation of the electrophoresis solution, improves the homogeneity between replicate samples in the 96-minigel format. The high-throughput method described in this paper should greatly increase the overall capacity, versatility and robustness of the comet assay.

journal_name

Mutagenesis

journal_title

Mutagenesis

authors

Gutzkow KB,Langleite TM,Meier S,Graupner A,Collins AR,Brunborg G

doi

10.1093/mutage/get012

subject

Has Abstract

pub_date

2013-05-01 00:00:00

pages

333-40

issue

3

eissn

0267-8357

issn

1464-3804

pii

get012

journal_volume

28

pub_type

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