Strand displacement detection of target nucleic acid
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United States of America Patent
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Issued Date -
Aug 14, 2003
app pub date -
Dec 7, 2001
filing date -
Dec 19, 2000
priority date (Note) -
Abandoned
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Abstract
Methods are provided for the determination of specific nucleotide sequences, particularly single nucleotide polymorphisms, by using probes comprising a first strand complementary to the target sequence, a shorter second strand forming a stem and a linker adjacent the double stranded stem connecting the two strands, whereby binding of target to the probe under conditions that do not cause melting of the double stranded stem in the absence of target results in the dissociation of the second strand from the first strand. The ss second strand is then detected as exemplified by using a FRET pair, where dissociation of the stem results in separation of the FRET pair and increase in fluorescence. Amplification of the target sequence may be employed prior to combination with the probe. The method finds particular application with complex nucleic acid mixtures.
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