Enzyme mixture

摘要

Polymorphisms are present throughout an organism's genome, and understanding which alleles are present in a particular organism's genome can be advantageous. When probing the identity of these alleles, one must minimize incorrect readings due to inefficiencies in the system. In hydrolysis probe applications, these inefficiencies may be due to over-activity of an exonuclease functionality that excises nucleotides from probes that are only partially, complementary to a region of a target. The present invention provides a mixture that contains a plurality of polymerases including one that has a 5′→3′ exonuclease functionality and one that lacks or substantially lacks it, each in a sufficient relative amount and concentration to increase efficiencies of the system.

主权项

1. A composition comprising
a first DNA polymerase, wherein the first polymerase has a 5′→3′ exonuclease activity; and a second DNA polymerase, wherein the second polymerase lacks or substantially lacks a 5′→3′ exonuclease activity, and at least one hydrolysis probe comprising a quencher moiety and a reporter moiety, and optionally a 3′ modification, the combined amount of first and second polymerases in the composition is between 100 polymerase activity units per milliliter and 500 polymerase activity units per milliliter, the ratio of the first polymerase to the second polymerase in the composition is 1:99 to 1:19.