US 9,809,843 B1
High throughput measurement of DNA base lesion repair capacity
Yuan Liu, Miami, FL (US); Yanhao Lai, Miami, FL (US); and Ruipeng Lei, Miami, FL (US)
Assigned to THE FLORIDA INTERNATIONAL UNIVERSITY BOARD OF TRUSTEES, Miami, FL (US)
Filed by Yuan Liu, Miami, FL (US); Yanhao Lai, Miami, FL (US); and Ruipeng Lei, Miami, FL (US)
Filed on Aug. 31, 2016, as Appl. No. 15/253,412.
Int. Cl. C12Q 1/68 (2006.01); G01N 21/64 (2006.01); G01N 21/29 (2006.01); G01N 33/68 (2006.01)
CPC C12Q 1/6827 (2013.01) [G01N 21/6428 (2013.01); G01N 2021/6432 (2013.01); G01N 2021/6439 (2013.01); G01N 2201/062 (2013.01)] 19 Claims
 
1. A method for measuring DNA repair capacity of a test solution, the method comprising the steps of:
a) incubating
i) a test reaction containing a damaged DNA comprising:
a marker attached to a first strand of the damaged DNA and
a quencher attached to a second strand of the damaged DNA, wherein the quencher inhibits the release of a detectable signal from the marker when the damaged DNA is in a double-stranded form, and the test solution, and optionally,
ii) a control reaction containing the damaged DNA and a control solution,
under conditions allowing the constituents of the test solution and the control solution to repair the damaged DNA such that the repaired DNA is double-stranded and the quencher inhibits the release of a detectable signal from the marker in the repaired DNA;
b) detecting a signal emitted from the marker released from the damaged DNA if the damaged DNA is not repaired;
c) measuring the amount of damaged DNA remaining not repaired in each of: i) the test reaction, and, if performed ii) the control reaction; and
d) determining the DNA repair capacity of the test solution based on:
i) the amount of damaged DNA remaining not repaired in the test reaction after the incubation of step a) compared to the amount of damaged DNA added in the test reaction, and/or
ii) the amount of damaged DNA remaining not repaired in the test reaction after the incubation of step a) compared to the amount of damaged DNA remaining not repaired in the control reaction after the incubation of step a).