| 摘要 |
The invention provides novel compounds and compositions of Formulas I and II, as well as methods of using them. The compounds can be used, for example, to quantify an amount of double stranded DNA in a sample subjected to nucleic acid amplification, or for real time monitoring of a nucleic acid amplification reaction. The compounds can be provided in a kit, for example, with other reagents and instructions for using the compounds and reagents. |
| 主权项 |
1. A method to quantify the amount of a nucleic acid in a sample subjected to nucleic acid amplification, comprising:
a) contacting the sample with a compound according to Formula I: wherein R1, R2 and R3 are each independently H, alkyl, cycloalkyl, aryl, heteroaryl, amino, hydroxy, or halo; or R1 and R2, or R2 and R3, taken together with the atoms to which they are attached form a fused benzo ring that is optionally substituted with 1, 2, 3, or 4 alkyl, cycloalkyl, aryl, heteroaryl, amino, hydroxy, or halo groups; Y is S, O, CF2, or NR4; R4 is (C1-C8)alkyl, aryl, or (C1-C8)alkaryl; W taken together with the atoms to which it is attached is a 5-, 6-, 7-, or 8-membered heterocyclic ring; n is 0, 1, or 2; Q is Q1 or Q2: wherein Y is —CR13═CR14—; m and p are 0 or 1 such that m+p=1; R6 is (C1-C8)alkyl, aryl, or (C1-C8)alkaryl; R7, R8, R9, R10, R11, and R12 are each independently H, alkyl, cycloalkyl, aryl, heteroaryl, amino, or halo; one of R13 and R14 is H and the other is NR15R16; R15 is (C1-C6)alkyl or (C1-C6)alkyl-NR17R18, wherein R17 and R18 are each independently H or (C1-C6)alkyl; and R16 is (C1-C6)alkyl-NR12R18, wherein R12 and R18 are each independently H or (C1-C6)alkyl; or a salt thereof; b) detecting fluorescence in the amplification reaction; and c) correlating the amount of fluorescence in the amplification reaction with the amount of nucleic acid in the reaction. |
