Albert M Bobst

US Patent:

7125665, Oct 24, 2006

Filed:

Jan 26, 2001

Appl. No.:

10/182559

Inventors:

Albert M. Bobst - Cincinnati OH, US
Jeffery D. Hester - Cincinnati OH, US

International Classification:

C12Q 1/68

US Classification:

435 6

Abstract:

The present invention relates to methods and kits for detecting the presence or absence of a target DNA sequence, such as a mutation, within an identified region of a selected DNA molecule, such as a gene. In particular aspects, the invention relates to the use of certain aspects of the polymerase chain reaction (“PCR”) and ligase chain reaction (“LCR”) techniques for the detection of genetic mutations in genes, particularly point mutations. A kit has been developed for direct EPR detection of specific PCR amplified target nucleic acid sequences. The PCR reaction is carried out in the presence of nitroxide-labeled oligomers that are degraded only if hybridized to a complementary target sequence. The degradation of the nitroxide-labeled oligomers into nitroxide-labeled cleavage products results in a characteristic increase of the h-/ho ratio of the EPR signal; in the absence of a complementary target sequence the EPR signal of nitroxide-labeled oligomer remains unchanged.

US Patent:

50048090, Apr 2, 1991

Filed:

Feb 4, 1986

Appl. No.:

6/826064

Inventors:

Albert M. Bobst - Cincinnati OH
Gary T. Pauley - Cincinnati OH

Assignee:

University of Cincinnati - Cincinnati OH

International Classification:

C07H 1700

US Classification:

536 29

Abstract:

Nitroxide labeled hybridization probes are formed from novel nucleotides. The novel nucleotides are formed by substituting a nitroxide moiety at the C-5 position of the pyrimidine ring of either cytosine or uracil. The hybridization probes are formed by nick translation with E. coli polymerase I or T4 DNA polymerase or other template dependent enzymes. The hybridization probes can then be detected using electron spin resonance spectroscopy or alternately using a colorimetric method.