Abstract: Provided are mutant polymerases having DNA polymerase activity and reverse transcriptase activity or strand displacement activity, along with target nucleic acid amplification methods employing such mutant polymerases.

Abstract: Provided are mutant polymerases having DNA polymerase activity and reverse transcriptase activity or strand displacement activity, along with target nucleic acid amplification methods employing such mutant polymerases.

Abstract: Provided are mutant polymerases having DNA polymerase activity and reverse transcriptase activity or strand displacement activity, along with target nucleic acid amplification methods employing such mutant polymerases.

Abstract: Provided are mutant polymerases having DNA polymerase activity and reverse transcriptase activity or strand displacement activity, along with target nucleic acid amplification methods employing such mutant polymerases.

Abstract: Provided are mutant polymerases having DNA polymerase activity and reverse transcriptase activity or strand displacement activity, along with target nucleic acid amplification methods employing such mutant polymerases.

Abstract: Provided herein is a method for homologous end cloning using a 5?-exonuclease. One aspect provides a method of generating a recombinant DNA molecule comprising 5? exonuclease digestion of DNA molecules, exonuclease inactivation, annealing of 3? overhangs to form a annealed complex. Also provided is a method a method of generating a recombinant DNA molecule comprising 5? exonuclease digestion of DNA molecules, exonuclease inactivation, annealing of 3? overhangs to form a annealed complex, and extension of 3? ends to fill gaps corresponding to exonuclease removal to form a substantially non-gapped annealed complex. Also provided is a one reaction method in which no further additions need be made to the reaction mixture and the reaction mixture is only manipulated thereafter as to incubation time and incubation temperature. Complexes formed according to methods described herein can be transformed into a host cell without further in vitro processing.

Abstract: An isolated polypeptide having fast elongating polymerase activity. Also provided are kits containing the isolated polypeptide and isolated polynucleotides encoding the isolated polypeptide.

Abstract: Provided herein is a method for homologous end cloning using a 5?-exonuclease. One aspect provides a method of generating a recombinant DNA molecule comprising 5? exonuclease digestion of DNA molecules, exonuclease inactivation, annealing of 3? overhangs to form a annealed complex. Also provided is a method a method of generating a recombinant DNA molecule comprising 5? exonuclease digestion of DNA molecules, exonuclease inactivation, annealing of 3? overhangs to form a annealed complex, and extension of 3? ends to fill gaps corresponding to exonuclease removal to form a substantially non-gapped annealed complex. Also provided is a one reaction method in which no further additions need be made to the reaction mixture and the reaction mixture is only manipulated thereafter as to incubation time and incubation temperature. Complexes formed according to methods described herein can be transformed into a host cell without further in vitro processing.

Abstract: A formulation and kit of thermostable or other DNA polymerases comprising at least one thermostable or other DNA polymerase which lacks 3?-exonuclease activity, and at least one thermostable DNA polymerase exhibiting 3?-exonuclease activity. Also provided is an improved method for enzymatic extension of DNA strands, especially while, but not limited to, amplifying nucleic acid sequences by polymerase chain reaction wherein the above formulation is made and used to catalyze primer extension.

Abstract: A formulation and kit of thermostable or other DNA polymerases comprising at least one thermostable or other DNA polymerase which lacks 3?-exonuclease activity, and at least one thermostable DNA polymerase exhibiting 3?-exonuclease activity. Also provided is an improved method for enzymatic extension of DNA strands, especially while, but not limited to, amplifying nucleic acid sequences by polymerase chain reaction wherein the above formulation is made and used to catalyze primer extension.

Abstract: A formulation of thermostable or other DNA polymerases comprising (a) at least one thermostable or other DNA polymerase which lacks 3?-exonuclease activity, and (b) at least one thermostable DNA polymerase exhibiting 3?-exonuclease activity, wherein the ratio of (a) to (b) is greater than 1 to 1. Also provided is an improved method for enzymatic extension of DNA strands, especially while, but not limited to, amplifying nucleic acid sequences by polymerase chain reaction wherein the above formulation is made and used to catalyze primer extension, are also provided.

Abstract: A method of obtaining DNA amplification of a nucleic acid target from a volume of whole blood comprising performing DNA amplification in a PCR assay mixture with a blood-resistant polymerase.

Abstract: A formulation and kit of thermostable or other DNA polymerases comprising at least one thermostable or other DNA polymerase which lacks 3?-exonuclease activity, and at least one thermostable DNA polymerase exhibiting 3?-exonuclease activity. Also provided is an improved method for enzymatic extension of DNA strands, especially while, but not limited to, amplifying nucleic acid sequences by polymerase chain reaction wherein the above formulation is made and used to catalyze primer extension.

Abstract: The present invention provides a method of performing hot start PCR reactions. The method is based on sequestration of magnesium ions in the form of a precipitate which renders a DNA polymerase inactive until the appropriate time in the PCR reaction when a certain temperature is reached and the magnesium ions are released from the precipitate. Also provided are kits comprising reagents and instructions for amplifying a target nucleic acid by the magnesium precipitate hot start.

Abstract: A formulation and kit of thermostable or other DNA polymerases comprising at least one thermostable or other DNA polymerase which lacks 3′-exonuclease activity, and at least one thermostable DNA polymerase exhibiting 3′-exonuclease activity. Also provided is an improved method for enzymatic extension of DNA strands, especially while, but not limited to, amplifying nucleic acid sequences by polymerase chain reaction wherein the above formulation is made and used to catalyze primer extension.

Abstract: The present invention provides methods of performing enzymatic reactions which require the use of magnesium dependent enzymes, including restriction endonucleases, ligases, and reverse transcriptases. The method is based on sequestration of magnesium ions in the form of a precipitate which renders a magnesium dependent enzyme inactive until the appropriate time in the reaction when a certain temperature is reached and the magnesium ions are released from the precipitate. Also provided are kits comprising reagents and instructions for DNA digestion and ligation, and for reverse transcription of RNA into cDNA. Furthermore, the kits and reagents of the present invention can be utilized in other reactions requiring magnesium dependent enzymes.

Abstract: A formulation and kit of thermostable or other DNA polymerases comprising at least one thermostable or other DNA polymerase which lacks 3′-exonuclease activity, and at least one thermostable DNA polymerase exhibiting 3′-exonuclease activity. Also provided is an improved method for enzymatic extension of DNA strands, especially while, but not limited to, amplifying nucleic acid sequences by polymerase chain reaction wherein the above formulation is made and used to catalyze primer extension.

Abstract: The present invention provides a method of performing hot start PCR reactions. The method is based on sequestration of magnesium ions in the form of a precipitate which renders a DNA polymerase inactive until the appropriate time in the PCR reaction when a certain temperature is reached and the magnesium ions are released from the precipitate. Also provided are kits comprising reagents and instructions for amplifying a target nucleic acid by the magnesium precipitate hot start.

Abstract: Provided are mutant DNA polymerases having at least one mutation which exhibit substantially reduced polymerase activity at 25° C. when compared to the same DNA polymerases without the at least one mutation and which exhibit normal or near-normal polymerase activity at optimum temperatures when compared to the same DNA polymerases without the at least one mutation. Also provided are amino acid sequences and nucleic acid sequences encoding such DNA polymerases, and vector plasmids and host cells suitable for the expression of these sequences. Also described herein are improved methods for performing polymerase chain reaction (PCR) amplification and other genetic manipulations and analyses using the mutant DNA polymerases of the invention.

Abstract: Provided are mutant DNA polymerases having at least one mutation which exhibit substantially reduced polymerase activity at 25° C. when compared to the same DNA polymerases without the at least one mutation and which exhibit normal or near-normal polymerase activity at optimum temperatures when compared to the same DNA polymerases without the at least one mutation. Also provided are amino acid sequences and nucleic acid sequences encoding such DNA polymerases, and vector plasmids and host cells suitable for the expression of these sequences. Also described herein are improved methods for performing polymerase chain reaction (PCR) amplification and other genetic manipulations and analyses using the mutant DNA polymerases of the invention.