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.