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Title: DNA polymerase having modified nucleotide binding site for DNA sequencing

Abstract

A modified gene encoding a modified DNA polymerase is disclosed. The modified polymerase incorporates dideoxynucleotides at least 20-fold better compared to the corresponding deoxynucleotides as compared with the corresponding naturally-occurring DNA polymerase. 6 figs.

Inventors:
;
Publication Date:
Research Org.:
Harvard University
OSTI Identifier:
458581
Patent Number(s):
US 5,614,365/A/
Application Number:
PAN: 8-337,615
Assignee:
Harvard Coll., Cambridge, MA (United States) PTO; SCA: 550200; PA: EDB-97:057520; SN: 97001758981
DOE Contract Number:
FG02-88ER60688
Resource Type:
Patent
Resource Relation:
Other Information: PBD: 25 Mar 1997
Country of Publication:
United States
Language:
English
Subject:
55 BIOLOGY AND MEDICINE, BASIC STUDIES; DNA POLYMERASES; GENES; DNA SEQUENCING; RECEPTORS; NUCLEOTIDES; POLYMERASE CHAIN REACTION; BIOTECHNOLOGY

Citation Formats

Tabor, S., and Richardson, C.. DNA polymerase having modified nucleotide binding site for DNA sequencing. United States: N. p., 1997. Web.
Tabor, S., & Richardson, C.. DNA polymerase having modified nucleotide binding site for DNA sequencing. United States.
Tabor, S., and Richardson, C.. 1997. "DNA polymerase having modified nucleotide binding site for DNA sequencing". United States. doi:.
@article{osti_458581,
title = {DNA polymerase having modified nucleotide binding site for DNA sequencing},
author = {Tabor, S. and Richardson, C.},
abstractNote = {A modified gene encoding a modified DNA polymerase is disclosed. The modified polymerase incorporates dideoxynucleotides at least 20-fold better compared to the corresponding deoxynucleotides as compared with the corresponding naturally-occurring DNA polymerase. 6 figs.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = 1997,
month = 3
}
  • Modified gene encoding a modified DNA polymerase wherein the modified polymerase incorporates dideoxynucleotides at least 20-fold better compared to the corresponding deoxynucleotides as compared with the corresponding naturally-occurring DNA polymerase.
  • SP6 bacteriophage RNA polymerase is produced by cultivating a new microorganism (particularly new strains of Escherichia coli) harboring a plasmid that carries SP6 bacteriophage RNA polymerase gene and recovering SP6 bacteriophage RNA polymerase from the culture broth. SP6 bacteriophage RNA polymerase gene is provided as are new microorganisms harboring a plasmid that carries SP6 bacteriophage RNA polymerase gene.
  • A method for rapid and efficient detection of a target DNA or RNA sequence is provided. A primer having a 3'-hydroxyl group at one end and having a sequence of nucleotides sufficiently homologous with an identifying sequence of nucleotides in the target DNA is selected. The primer is hybridized to the identifying sequence of nucleotides on the DNA or RNA sequence and a reporter molecule is synthesized on the target sequence by progressively binding complementary nucleotides to the primer, where the complementary nucleotides include nucleotides labeled with a fluorophore. Fluorescence emitted by fluorophores on single reporter molecules is detected tomore » identify the target DNA or RNA sequence.« less
  • Compositions and methods for making a plurality of probes for analyzing a plurality of nucleic acid samples are provided. Compositions and methods for analyzing a plurality of nucleic acid samples to obtain sequence information in each nucleic acid sample are also provided.
  • This patent describes a method for determining the base sequence of nucleotides. It comprises: adding an activated nucleoside 5'-triphosphate precursor of one known nitrogenous base composition to a reaction mixture comprising a template-directed nucleotide polymerase and a single-stranded polynucleotide templates hybridized to complementary oligonucleotide primer strands at least one nucleotide residue shorter than the templates to form at least one unpaired nucleotide residue in each template at the 3'-end of the primer strand under reaction conditions which allow incorporation of the activated nucleoside 5'-triphosphate precursor onto the 3' end of the primer strands; detecting whether or not the nucleoside 5'triphosphatemore » precursor was incorporated into the primer strands; sequentially repeating these steps; and determining the base sequence of the unpaired nucleotide residues of the template from the sequence of incorporation of the nucleoside precursors.« less