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A FLUORESCENCE BASED ASSAY FOR DNA DAMAGE INDUCED BY STYRENE OXIDE
Citation:
Ramanathan, K AND K R. Rogers. A FLUORESCENCE BASED ASSAY FOR DNA DAMAGE INDUCED BY STYRENE OXIDE. SENSORS & ACTUATORS B 91(1-3):205-210, (2003).
Impact/Purpose:
The overall objective of this task is to develop rapid, cost-effective and scientifically sound techniques for measuring chemically induced DNA damage. This method is expected to provide the Agency with rapid, sensitive, and simple techniques that can be used among a panel of methods to determine the genotoxic potential of polluted samples.
Description:
A rapid and simple assay to detect DNA damage to calf thymus DNA caused by styrene oxide (SO) is reported. This assay is based on changes observed in the melting and annealing behavior of the damaged DNA. The melting annealing process was monitored using a fluorescence indicator dye PicoGreen (R) (PG) that increases its fluorescence when bound to double stranded DNA. The assay response was monitored with varying concentrations of SO, incubation temperature, incubation time and salt concentration. Sample volumes of 10 ul containing 100 ng/ml DNA were required for this assay. The error in measurement was between 5 and 7%.
The U.S. Environmental Protection Agency (EPA), through its Office of Research and Development (ORD), has funded this research. It has been subject to the EPA's peer and administrative review and approved for publication. Mention of trade names or commercial products does not constitute endorsement or recommendation by EPA for use.