Method of using T7 DNA polymerase to label the 3' end of a DNA molecule

• US 5,266,466 A
• Filed: 09/04/1992
• Issued: 11/30/1993
• Est. Priority Date: 01/14/1987
• Status: Expired due to Term

• Current Assignee: President and Fellows of Harvard College (Harvard Corporation)
• Original Assignee: President and Fellows of Harvard College (Harvard Corporation)
• Inventors: Tabor, Stanley, Richardson, Charles C.
• Primary Examiner(s): Schwartz, Richard A.
• Assistant Examiner(s): HODGES, ROBERT
• Application Number:
  US07/940,531
• Time in Patent Office: 452 Days
• Field of Search: 435/5, 435/6, 435/91, 435/172.3, 435/172.1, 935/17, 935/77
• US Class Current: 435/91.5
• 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 1/6858 Allele-specific amplification
• CPC Class Codes: C12Q 1/686 Polymerase chain reaction [PCR]
• CPC Class Codes: C12Q 1/6869 Methods for sequencing
• CPC Class Codes: C12Q 2521/101 DNA polymerase
• CPC Class Codes: C12Q 2535/101 Sanger sequencing method, i.e. oligonucleotide sequencing using primer elongation and dideoxynucleotides as chain terminators

Abstract

This invention relates to T7-type DNA polymerases and methods for using them.

Claims

1. A method for labelling the 3'"'"' end of a DNA fragment comprising incubating said DNA fragment with T7-type DNA polymerase produced from recombinant DNA, having less than 500 units of exonuclease activity per mg of polymerase, and a labelled deoxynucleotide species under conditions in which said 3'"'"' end of said DNA fragment is extended by said polymerase and thereby labelled by addition of said labelled deoxynucleotide to said DNA fragment.