engleski

Assessing the Fast Micromethod as a tool in DNA integrity measurement

Over the past decade the relatively new Fast Micromethod® for detecting DNA damage (strand breaks, alkali-labile sites and incomplete excision repair) and related DNA integrity in cell suspensions or tissue homogenates in single microplates has been developed. The technique is based on the ability of the specific fluorophore PicoGreen® to preferentially interact with high integrity DNA molecules, dsDNA, allowing direct fluorimetric measurement of fluorophore-dsDNA complex decay in alkaline conditions. This analytical technique allows simple and fast simultaneous analysis of DNA integrity in numerous, low DNA content samples. Research has confirmed that the Fast Micromethod® is applicable for the measurement of DNA integrity in not only human biological material for medical purpose, but also in marine invertebrates for genotoxicity assessment (biomonitoring) and the estimation of harmful effects in the ecosystem. This article focuses on a comprehensive description of the method and its uses, and offers detailed descriptions and insights into samples and chemicals preparation, fluorescence analysis and calculation of DNA integrity rate. The results and features of many provided studies using this method are discussed, in order to elucidate advantages and imperfections of this analytical technique and to highlight approaches that may allow its further development.

DNA alkali-labile sites, DNA cross-links, DNA integrity, DNA strand breaks, Fast Micromethod®

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