AN.012.S60
The use of fluorescence stopped-flow methods for the elucidation of rapid kinetics in solution is well characterised. By using a plane polarised excitation beam, however, the intensity of fluorescence in directions parallel and perpendicular to the direction of polarisation can be measured. These two components can be combined so as to express a convenient technique to investigate the binding of a protein to DNA. This technique uses an oligonucleotide that is labelled during synthesis with a commercially available fluorescent dye. It can thus be applied to any DNA binding protein and does not rely on any intrinsic fluorescence properties of the protein. This application note reports on the binding of a DNA repair enzyme, uracil DNA glycosylase, to an oligonucleotide containing uracil.