Polypeptides Having Nucleic Acid Binding Activity and Compositions and Methods For Nucleic Acid Amplification
0 Assignments
0 Petitions
Accused Products
Abstract
Polypeptides having nucleic acid binding activity are provided. Methods of using polypeptides having nucleic acid binding activity are provided. Fusion proteins and methods of using fusion proteins are provided. Fusion proteins comprising a polymerase and a nucleic acid binding polypeptide are provided. Fusion proteins comprising a reverse transcriptase and a nucleic acid binding polypeptide are provided. Methods are provided for amplifying a nucleic acid sequence using a fusion protein comprising a nucleic acid binding polypeptide and a polymerase. Methods are provided for amplifying a nucleic acid sequence using a fusion protein comprising a nucleic acid binding polypeptide and a reverse transcriptase.
92 Citations
120 Claims
-
1-90. -90. (canceled)
-
91. A method of amplifying a nucleic acid sequence, wherein the method comprises subjecting a reaction mixture to at least one amplification cycle, wherein the reaction mixture comprises a double-stranded nucleic acid, at least two primers which anneal to complementary strands of the double-stranded nucleic acid, and a fusion protein comprising a thermostable DNA polymerase and a nucleic acid binding polypeptide, and wherein the at least one amplification cycle comprises:
-
a) denaturing the double-stranded nucleic acid; b) annealing the at least two primers to complementary strands of the denatured double-stranded nucleic acid; and c) extending the at least two primers for an extension time to obtain primer extension products; and wherein the time to complete one amplification cycle is 20 seconds or less and wherein the extension time is between 2 to 10 seconds per thousand base pairs of extension products. - View Dependent Claims (92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118)
-
-
119. A method of generating DNA from RNA template comprising exposing the RNA template to at least one primer and a fusion protein comprising a nucleic acid binding polypeptide and a polymerase, wherein the polymerase is a family B polymerase, a fragment of a family B polymerase, or a polypeptide having at least 80% identity to a family of B polymerases, wherein the fusion protein has reverse transcriptase activity.
-
120. A fusion protein comprising:
-
a) a polypeptide comprising an amino acid sequence of a nucleic acid binding polypeptide or a fragment thereof having nucleic acid binding activity; and b) a reverse transcriptase.
-
Specification