Polymerase enhancing factor (PEF) extracts, PEF protein complexes, isolated PEF protein, and methods for purifying and identifying
AssigneeStratagene
InventorsHolly Hogrefe, Connie Hansen
Life of this patent
Estimating the term…
Abstract
The invention provides novel extracts, proteins, and complexes that improve the polymerization activity of nucleic acid polymerases. Included within the aspects of the invention are methods for identifying compositions with a polymerase enhancing activity, methods for purifying and using these compositions, and specific extracts, proteins, and complexes that function to enhance polymerase activity. As an example, specifically described is nucleotide and amino acid sequence information for a Pyrococcus furiousus PEF (P45), which was used to produce a recombinant PEF.
Claims18
1-95. (canceled)
A composition comprising:
- (a) an enzyme possessing substantial 3′ to 5′ exonuclease activity, (b) a DNA polymerase, wherein the DNA polymerase has less 3′ to 5′ exonuclease activity than the enzyme, and (c) a factor that substantially inhibits the incorporation of undesired nucleotides or analogs thereof into a DNA polymer.
A method for amplifying at least one nucleic acid comprising subjecting an amplification reaction composition comprising:
- (a) an enzyme possessing substantial 3′ to 5′ exonuclease activity, (b) a DNA polymerase, wherein the DNA polymerase has less 3′ to 5′ exonuclease activity than the enzyme, (c) a factor that substantially inhibits the incorporation of undesired nucleotides or analogs thereof into a DNA polymer, and (d) at least one nucleic acid to at least one cycle of amplification to amplify the at least one nucleic acid.
A kit for amplifying, synthesizing, or mutagenizing nucleic acids comprising:
- (a) an enzyme possessing substantial 3′ to 5′ exonuclease activity, (b) a DNA polymerase, wherein the DNA polymerase has less 3′ to 5′ exonuclease activity than the enzyme, and (c) a factor that substantially inhibits the incorporation of undesired nucleotides or analogs thereof into a DNA polymer.
Dependencies are inferred from the claim text.