.phi.29 DNA polymerase

Details .phi.29 DNA polymerase .phi.29 DNA polymerase

1993-02-11

1996-11-19

Zitomer, Stephanie W.

Chemistry: molecular biology and microbiology

Enzyme , proenzyme; compositions thereof; process for...

Transferase other than ribonuclease

C12N 912

Patent

active

055762042

ABSTRACT:
An improved method for determining the nucleotide base sequence of a DNA molecule employs a .phi.-29 type DNA polymerase modified to have reduced or no exonuclease activity. The method includes annealing the DNA molecule with a primer molecule able to hybridize to the DNA molecule; incubating the annealed mixture in a vessel containing four different deoxynucleoside triphosphates, a DNA polymerase, and one or more DNA synthesis terminating agents which terminate DNA synthesis at a specific nucleotide base, wherein each the agent terminates DNA synthesis at a different nucleotide base; and separating the DNA products of the incubating reaction according to size, whereby at least a part of the nucleotide base sequence of the DNA can be determined. The improvement is provision of a DNA polymerase which is a .phi.29-type DNA polymerase.

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