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BURACZEWSKA
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Tytuł oryginału:
A modified neutral comet assay: elimination of lysis at high temperature and validation of the assay with anti-single-stranded DNA antibody.
Autorzy:
Wojewódzka
Maria,
Buraczewska
Iwona,
Kruszewski
Marcin
Źródło:
Mutat. Res 2002: 518 (1) s.9-20, il., bibliogr. 43 poz.
Sygnatura GBL:
304,952
Hasła klasyfikacyjne GBL:
genetyka
Typ dokumentu:
praca doświadczalna
praca opublikowana za granicą
tytuł obcojęzyczny
Streszczenie angielskie:
Comet assay under neutral conditions allows the detection of DNA double-strand breaks (DSB), considered to be the biologically relevant radiation-induced lesion. In this report, we describe modifications of the neutral comet method, which simplify and facilitate its use for estimation of DNA DSB in X-irradiation mammalian cells in centure. The analysis carried out according to this protocol takes less time than those most often applied. Also, the use of lysis at 50ř C is avoided; this is important in view of the presence of heat-labile sites in the chromatin of irradiated cells, recently reported by Rydberg [Radiation-induced heat-labile sites that convert into DNA double-strtand breaks, Radsaiation Research 153 (2000) 805-812]. The comets have well-defined, sharp limits, suitable for image analysis. The chromatin of the hydrogen peroxide-treated or UV-C-irradiated cell remains condensed similarly to that of the control cells. We checked the neutral comets for the presence of single-stranded DNA by means of a specific antibody. The results point to a satisfactory sensitivity of the modified neutral comet assay and its specificity for DSB. The minimum detection level of the modified neutral comet assay is about 5 Gy.
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