The comet assay as a method of identifying chromosomes instability

被引:7
|
作者
Czubaszek, Magdalena [1 ]
Szostek, Malgorzata [1 ]
Wojcik, Ewa [1 ]
Andraszek, Katarzyna [1 ]
机构
[1] Uniwersytet Przyrodniczohumanistyczny Siedlcach, Katedra Genetyki & Hodowli Koni, PL-08110 Siedlce, Poland
来源
POSTEPY HIGIENY I MEDYCYNY DOSWIADCZALNEJ | 2014年 / 68卷
关键词
DNA; chromosomal instability; chromosomal aberrations; comet assay; SISTER-CHROMATID EXCHANGES; INDUCED DNA-DAMAGE; CELLS; GENOTOXICITY; SPERMATOZOA; FRAGMENTATION; EXTENDERS; INTEGRITY; PROGRAM; STORAGE;
D O I
10.5604/17322693.1106384
中图分类号
R-3 [医学研究方法]; R3 [基础医学];
学科分类号
1001 ;
摘要
The basic method for analyzing the degree of DNA fragmentation caused by genotoxic factors is gel electrophoresis of single cells (single cell gel electrophoresis), also called the comet assay. The comet assay enables the analysis of the level of several different DNA modifications. The basic testing procedure has been only slightly modified. This method helps identify single-strand and double-strand DNA cracks, as well as any chemical and enzymatic modifications that can potentially turn into cracks in DNA or chromatids. The comet assay makes it possible to detect DNA damage at the level of single cells. It can be employed in analyses of any tissues which provide cellular suspensions. Analysed cells are submerged in agarose on a microscope slide. DNA is what is left after proteins have been broken down. The slide is then subjected to electrophoresis and stained with a fluorescent dye. A "comet-like" image is obtained. The "head" is the cell fixation site prior to lysis; the "tail" represents damaged DNA fragments. The extent of DNA damage is reflected in the length of the tail and the amount of DNA contained in it. The assay finds research applications in the following fields: genetic toxicology, monitoring of DNA repair following chemotherapy and radiotherapy, ecotoxicology, animal and human nourishment, biomonitoring of genotoxicity, epidemiology and assessment of material deposited in sperm and blood banks.
引用
收藏
页码:695 / 700
页数:6
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