G - Physics – 01 – N
Patent
G - Physics
01
N
195/1.23, 150/8.
G01N 33/52 (2006.01) C12Q 1/68 (2006.01)
Patent
CA 1161736
TITLE FLUORESCENT DETECTION OF DNA DAMAGE INVENTOR Hyman C. Birnboim ABSTRACT OF THE DISCLOSURE A sensitive method for detecting damage to DNA, e.g. resulting from exposure of living cells or whole organisms to low doses of radia- tion or chemicals, is described comprising: partial lysis of exposed cells to access the DNA; alkaline denaturing the DNA; after a selected interval stopping the denaturation by lowering the pH to a selected value; rendering the lysate homogeneous; adding an appropriate fluores- cent dye which interacts preferentially with double-stranded DNA and measuring the resulting fluorescence. The decrease in fluorescence (compared to that before denaturation) is a measure of the rate of DNA denaturation which is directly proportional to the extent of DNA damage. Several factors, particularly the lowered pH at which the denaturation is stopped, have been found important for increased sensitivity. The method is suitable for monitoring the effect on DNA in living cells of environ- mental factors, drug and radiation therapy, and for toxicology studies. A kit adapted for carrying out the test method is described.
393338
Her Majesty In Right Of Canada As Represented By Atomic Energy O
Thomson Alan A.
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