DNA polymerases having improved labeled nucleotide incorporation properties

• US 7,897,738 B2
• Filed: 12/09/2005
• Issued: 03/01/2011
• Est. Priority Date: 03/12/1997
• Status: Expired due to Term

• Current Assignee: Applied Biosystems LLC (Thermo Fisher Scientific Incorporated)
• Original Assignee: Applied Biosystems LLC (Thermo Fisher Scientific Incorporated)
• Inventors: Brandis, John, Richards, John H., Bloom, Curtis
• Primary Examiner(s): Chunduru; Suryaprabha
• Application Number:
  US11/299,124
• Publication Number:
  US 20060088879A1
• Time in Patent Office: 1,908 Days
• Field of Search: 530/350, 435/6, 435/91, 435/2, 536/23.1
• US Class Current: 536/23.1
• CPC Class Codes: C12N 9/1252 DNA-directed DNA polymerase (2.7.7.7), i.e. DNA replicase
• CPC Class Codes: C12P 19/34 Polynucleotides, e.g. nucleic acids, oligoribonucleotides
• CPC Class Codes: C12Q 1/6806 Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 1/6844 Nucleic acid amplification reactions
• CPC Class Codes: C12Q 1/6869 Methods for sequencing
• CPC Class Codes: C12Q 2521/101 DNA polymerase
• CPC Class Codes: C12Q 2563/107 fluorescence

Abstract

The present teachings relate to DNA polymerases that have an F667Y mutation as defined with respect to Taq DNA polymerase and exhibit reduced discrimination against labeled nucleotides into polynucleotides.

The teachings also include kits comprising the subject DNA polymerases and numerous methods of using the subject DNA polymerases, including uses for chain termination sequencing and PCR.

The present teachings relate to DNA polymerases that have an F667Y mutation as defined with respect to Taq DNA polymerase and exhibit reduced discrimination against labeled nucleotides into polynucleotides. The teachings also include kits comprising the subject DNA polymerases and numerous methods of using the subject DNA polymerases, including uses for chain termination sequencing and PCR.

Claims

1. A composition comprising a first DNA polymerase and a second DNA polymerase, wherein the first and the second DNA polymerase have an F667Y mutation as defined with respect to Taq DNA polymerase, and wherein the first DNA polymerase and the second DNA polymerase have primary amino acid sequences that differ from one another, and wherein the first DNA polymerase is derived from Taq DNA polymerase.