Appeal No. 1999-1266
Application No. 08/859,472
contacting a target nucleic acid sequence with PCR reagents, including at
least two PCR primers and a polymerase enzyme substantially lacking any 5'÷ 3'
exonuclease activity, and an oligonucleotide probe comprising:
an oligonucleotide;
a fluorescer molecule attached to a first end of the oligonucleotide;
a quencher molecule attached to a second end of the oligonucleotide such
that quencher molecule substantially quenches the fluorescer of the fluorescer molecule
whatever the oligonucleotide probe is in a single-stranded state and such that the
fluorescer is substantially unquenched whenever the oligonucleotide probe is in a double-
stranded state; and
a 3' end which is rendered impervious to the 5'÷ 3' extension activity of a
polymerase; and
subjecting the target nucleic sequence, the oligonucleotide probe, and the
PCR reagents to thermal cycling sufficient to amplify the target nucleic acid sequence
specified by the PCR reagents.
The references relied upon by the examiner are:
Abramson et al. (Abramson) 5,466,591 Nov. 14, 1995
Link et al. (Link) 9310267 May. 27, 1993
(WO)
Heller et al. (Heller) 0 229 943 Jul. 29, 1987
(European Patent)
Parkhurst et al. (Parkhurst) “Kinetic Studies by Fluorescence Energy Transfer Employing a
Double-Labeled Oligonucleotide: Hybridization to the Oligonucleotide Complement and to
Single-Stranded DNA,” Biochemistry, Vol. 34, pp. 285-292 (1995)
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