Polymerase inhibitors and related compositions and methods
The present disclosure includes compositions and methods for improved DNA amplification reactions. In particular, the present disclosure provides compositions and methods for hot-start PCR applications using DNA polymerase inhibitors that minimize non-specific DNA amplification by inactivating DNA polymerase at lower temperatures.
1. A composition comprising a compound of formula (II):
or a salt thereof,
wherein:
A is selected from a monocyclic or bicyclic aryl, heteroaryl, or heterocyclyl group, each of which may be optionally substituted with 1, 2, or 3 substituents;
R 1 is C 6 -C 20 alkyl;
R 2 is selected from hydrogen and —COOH;
n is 1 or 2; and
R 3 is selected from —COOH and —SO 3 X, wherein X is selected from hydrogen, an alkali metal cation, and an ammonium cation; and
one or more nucleic acid amplification reagents.
2. The composition of claim 1 , wherein A is phenyl that is unsubstituted or substituted with 1, 2, 3, or 4 substituents independently selected from C 1 -C 4 alkyl, C 1 -C 4 alkoxy, and halo.
3. The composition of claim 1 , wherein A is a 5- or 6-membered monocyclic heteroaryl that is unsubstituted or substituted with 1, 2, or 3 substituents independently selected from C 1 -C 4 alkyl, C 1 -C 4 alkoxy, and halo.
4. The composition of claim 1 , wherein A is a bicyclic heterocyclyl group that is unsubstituted.
5. The composition of claim 4 , wherein A is selected from 2,3-dihydrobenzofuranyl and chromanyl.
6. The composition of claim 1 , wherein R 1 is C 8 -C 14 alkyl.
7. The composition of claim 6 , wherein R 1 is C 12 alkyl.
8. The composition of claim 1 , wherein R 2 is hydrogen.
9. The composition of claim 1 , wherein R 2 is —COOH.
10. The composition of claim 1 , wherein n is 1.
11. The composition of claim 1 , wherein n is 2.
12. The composition of claim 1 , or a salt thereof, wherein R 3 is —COOH.
13. The composition of claim 1 , or a salt thereof, wherein R 3 is —SO 3 X, and X is a sodium cation.
14. The composition of claim 1 , wherein the compound is in the form of an alkali metal salt.
15. The composition of claim 1 , wherein the compound of formula (II) is selected from:
16. The composition of claim 1 , wherein the one or more amplification reagents are selected from the group consisting of: a polymerase, deoxynucleotide triphosphates, buffer, a magnesium salt, an oligonucleotide primer, and a nucleic acid template.