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Method of using a TAQ DNA polymerase without 5'-3'-exonuclease activity

  • US 5,108,892 A
  • Filed: 08/03/1989
  • Issued: 04/28/1992
  • Est. Priority Date: 08/03/1989
  • Status: Expired due to Term
First Claim
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1. A method for determining the nucleotide base sequence of a DNA molecule, comprising:

  • a) hybridizing a primer to a DNA template molecule to be sequenced;

    b) extending the primer with a Taq DNA polymerase having an apparent molecular weight of about 80,000 daltons and substantially no 5'"'"'-3'"'"' exonuclease activity;

    c) incorporating a strand terminating nucleotide onto the extended primer under conditions to yield a distinct population of nucleotide fragments beginning on the same nucleotide position for all populations and having a variable terminus at one of the following four nucleotide bases;

    A, C, G and T; and

    d) separating the synthesized fragments according to their size, whereby at least a part of the nucleotide base sequence of the DNA molecule can be determined.

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