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Quantitative method for the evaluation of the global capabilities and specific for the repair of the dna of at least one biological medium, as well as its applications
Quantitative method for the evaluation of the global capabilities and specific for the repair of the dna of at least one biological medium, as well as its applications
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机译:评估至少一种生物介质的整体能力和修复DNA的能力的定量方法及其应用
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摘要
Quantitatively evaluating the global and specific capacities for DNA repair in a biological medium, is new. Process for quantitative evaluation of the global and specific capacities for DNA repair in a biological medium comprises: (a) preparing a series of plasmids (P), each containing distinct DNA lesions, by independently treating them with a physical and/or chemical agent and recovery of the supercoiled fraction; (b) characterizing the lesions in each P; (c) depositing P, and at least one control supercoiled plasmid without lesions, on a single solid support, according to a pre-established scheme, to produce a functional support divided into x distinct zones (x = number of media to be tested simultaneously) where each zone contains the set of P; (d) incubating the support with different repair solutions, each containing the enzyme activities required for repair, ATP (adenosine triphosphate) and a system for regenerating it, a labeled nucleotide triphosphate and other components required for enzymatic activity, at 30 degrees Centigrade for 1-5, preferably 3, hours, with each repair solution being applied to a different zone A1-Ax; (e) washing the support; (f) (in)direct measurement of the signal produced by the label, incorporated into DNA during the repair reaction, in each zone; (g) recording and quantifying the signal from each plasmid in each zone; and (h) determining the ratio of signal from each plasmid relative to that from the control.
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