Patent ID: 8021839

Claim:
A method of detecting a target polynucleotide in a sample, comprising the steps of: (a) producing a reaction mixture comprising the sample, a first polynucleotide, and a dye, wherein the first polynucleotide is at least partially complementary to a segment of the target polynucleotide, and wherein some of the dye forms a complex with the first polynucleotide and the target polynucleotide, if present in the sample; (b) exposing the reaction mixture to a light, wherein the light catalyzes a reaction that causes a decrease in the amount of the light reactive dye; and (c) observing the optical property of the reaction mixture at least once after exposure to the light and subsequent to the formation of the complex; wherein the reaction mixture has an optical property that changes in response to the light exposure if the first polynucleotide and the target polynucleotide are present therein, and wherein the dye is the compound of formula (I), or a salt or ester thereof: wherein, independently at each occurrence: R 1 and R 2 are each independently selected from hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, aryl, acyl, heteroacyl, heteroaryl, hydroxyl, alkoxy, carbonyl, sulfinyl, sulfonyl, and amino groups; R 3 is selected from the group consisting of hydrogen, alkyl, alkenyl, alkynyl, aryl, heteroalkyl, heteroalkenyl, heteroalkynyl, aryl, acyl, heteroacyl, heteroaryl, aryl, alkyl, heteroarylalkyl, hydroxyl, alkoxy, halo, carbonyl, sulfinyl, sulfonyl, and amino groups; R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 , R 12 , and R 13 are each independently selected from the group consisting of hydrogen, halogen, alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, aryl, acyl, heteroacyl, heteroaryl, aryl, alkyl, heteroarylalkyl, hydroxyl, alkoxy, carbonyl, sulfinyl, sulfonyl groups; n is 0, 1, 2, 3, 4, or 5; and Y is —CR 12 ═CR 13 —, sulfur, or oxygen.