Sensitive high throughput method for DNA damage and repair

摘要

A high throughput method and apparatus for rapidly screening a plurality of genotoxicants to determine the degree and type of genotoxicity are provided.

主权项

1. A high throughput method for rapidly screening a plurality of genotoxicants to simultaneously detect degree and type of DNA damage comprising the steps of:
(a) adding a volume of dsDNA to test wells; (b) adding a volume of genotoxicants to be evaluated to the test wells; (c) adding dsDNA specific dye and reference dye to the test wells; (d) monitoring a detectable signal, using fluorescent or spectrophotometric technique, emitted from test wells while melting the DNA; and (e) analyzing melting curves for the
(i) initial detectable signal,(ii) initial melting rate and(iii) temperature, pH or salinity level of initial melting,and comparing these parameters to those of controls so that degree and type of DNA damage caused by the genotoxicants can be simultaneously detected.