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Title: Kit for detecting nucleic acid sequences using competitive hybridization probes

Patent ·
OSTI ID:873914

A kit is provided for detecting a target nucleic acid sequence in a sample, the kit comprising: a first hybridization probe which includes a nucleic acid sequence that is sufficiently complementary to selectively hybridize to a first portion of the target sequence, the first hybridization probe including a first complexing agent for forming a binding pair with a second complexing agent; and a second hybridization probe which includes a nucleic acid sequence that is sufficiently complementary to selectively hybridize to a second portion of the target sequence to which the first hybridization probe does not selectively hybridize, the second hybridization probe including a detectable marker; a third hybridization probe which includes a nucleic acid sequence that is sufficiently complementary to selectively hybridize to a first portion of the target sequence, the third hybridization probe including the same detectable marker as the second hybridization probe; and a fourth hybridization probe which includes a nucleic acid sequence that is sufficiently complementary to selectively hybridize to a second portion of the target sequence to which the third hybridization probe does not selectively hybridize, the fourth hybridization probe including the first complexing agent for forming a binding pair with the second complexing agent; wherein the first and second hybridization probes are capable of simultaneously hybridizing to the target sequence and the third and fourth hybridization probes are capable of simultaneously hybridizing to the target sequence, the detectable marker is not present on the first or fourth hybridization probes and the first, second, third, and fourth hybridization probes each include a competitive nucleic acid sequence which is sufficiently complementary to a third portion of the target sequence that the competitive sequences of the first, second, third, and fourth hybridization probes compete with each other to hybridize to the third portion of the target sequence.

Research Organization:
Lawrence Livermore National Laboratory (LLNL), Livermore, CA (United States)
DOE Contract Number:
W-7405-ENG-48
Assignee:
Regents of University of California (Berkeley, CA)
Patent Number(s):
US 6270972
OSTI ID:
873914
Country of Publication:
United States
Language:
English

References (3)

Ligation of oligonucleotides to nucleic acids or proteins via disulfide bonds journal May 1988
Gene mapping and gene enrichment by the avidin-biotin interaction: use of cytochrome-c as a polyamine bridge journal January 1978
Enzymatic synthesis of biotin-labeled polynucleotides: novel nucleic acid affinity probes. journal November 1981