7 documents found, page 1 of 1
Inter-laboratory variation in measurement of DNA damage by the alkaline comet a...
Moller, P; Azqueta, A; Collia, M; Bakuradze, T; Richling, E; Bankoglu, EE; Stopper, H; Bastos, VC; Langie, SAS; Jensen, A; Ristori, S; Scavone, FThe comet assay is a simple and versatile method for measurement of DNA damage in eukaryotic cells. More specifically, the assay detects DNA migration from agarose gel-embedded nucleoids, which depends on assay conditions and the level of DNA damage. Certain steps in the comet assay procedure have substantial impact on the magnitude of DNA migration (e.g. electric potential and time of electrophoresis). Inter-l...
DNA strand break levels in cryopreserved mononuclear blood cell lines measured ...
Moller, P; Azqueta, A; Rodriguez-Garraus, A; Bakuradze, T; Richling, E; Bankoglu, EE; Stopper, H; Bastos, VC; Langie, SAS; Jensen, A; Ristori, SThe comet assay is widely used in biomonitoring studies for the analysis of DNA damage in leukocytes and peripheral blood mononuclear cells. Rather than processing blood samples directly, it can be desirable to cryopreserve whole blood or isolated cells for later analysis by the comet assay. However, this creates concern about artificial accumulation of DNA damage during cryopreservation. In this study, 10 labo...
Long-term cryopreservation of potassium bromate positive assay controls for mea...
Moller, P; Azqueta, A; Rodriguez-Garraus, A; Bakuradze, T; Richling, E; Bankoglu, EE; Stopper, H; Bastos, VC; Langie, SAS; Jensen, A; Ristori, SThe formamidopyrimidine DNA glycosylase (Fpg)-modified comet assay is widely used for the measurement of oxidatively generated damage to DNA. However, there has not been a recommended long-term positive control for this version of the comet assay. We have investigated potassium bromate as a positive control for the Fpg-modified comet assay because it generates many Fpg-sensitive sites with a little concurrent g...
Visual comet scoring revisited: a guide to scoring comet assay slides and obtai...
Moller, P; Azqueta, A; Sanz-Serrano, J; Bakuradze, T; Richling, E; Bankoglu, EE; Stopper, H; Bastos, VC; Langie, SAS; Jensen, A; Scavone, FMeasurement of DNA migration in the comet assay can be done by image analysis or visual scoring. The latter accounts for 20%-25% of the published comet assay results. Here we assess the intra- and inter-investigator variability in visual scoring of comets. We include three training sets of comet images, which can be used as reference for researchers who wish to use visual scoring of comets. Investigators in 11 ...
DNA damage in circulating leukocytes measured with the comet assay may predict ...
Bonassi, S; Ceppi, M; Møller, P; Azqueta, A; Milić, M; Neri, M; Brunborg, G; Godschalk, R; Koppen, G; Langie, SAS; Teixeira, JP; Bruzzone, MThe comet assay or single cell gel electrophoresis, is the most common method used to measure strand breaks and a variety of other DNA lesions in human populations. To estimate the risk of overall mortality, mortality by cause, and cancer incidence associated to DNA damage, a cohort of 2,403 healthy individuals (25,978 person-years) screened in 16 laboratories using the comet assay between 1996 and 2016 was fol...
Minimum Information for Reporting on the Comet Assay (MIRCA): recommendations f...
Møller, P; Azqueta, A; Boutet-Robinet, E; Koppen, G; Bonassi, S; Milić, M; Gajski, G; Costa, S; Teixeira, JP; Costa Pereira, C; Dusinska, MThe comet assay is a widely used test for the detection of DNA damage and repair activity. However, there are interlaboratory differences in reported levels of baseline and induced damage in the same experimental systems. These differences may be attributed to protocol differences, although it is difficult to identify the relevant conditions because detailed comet assay procedures are not always published. Here...
Potassium bromate as positive assay control for the Fpg-modified comet assay
Møller, P; Muruzabal, D; Bakuradze, T; Richling, E; Bankoglu, EE; Stopper, H; Langie, SAS; Azqueta, A; Jensen, A; Scavone, F; Giovannelli, LThe comet assay is a popular assay in biomonitoring studies. DNA strand breaks (or unspecific DNA lesions) are measured using the standard comet assay. Oxidative stress-generated DNA lesions can be measured by employing DNA repair enzymes to recognise oxidatively damaged DNA. Unfortunately, there has been a tendency to fail to report results from assay controls (or maybe even not to employ assay controls). We b...