Appeal No. 1999-1266
Application No. 08/859,472
the use of the specific fluorescent donor and quencher in double stranded
states. Link also does not teach the use of a 5'-3' exonuclease deficient
polymerase enzyme.
The examiner cites Parkhurst as teaching “the use of doubly labeled oligonucleotide
probes in which the probes are quenched in a single stranded state and unquenched in a
double stranded state (291, figure 3).” (Id.). The examiner, additionally, cites Heller as
teaching “the use of doubly labeled oligonucleotide probes in which the probes are quenched
in a single stranded state and unquenched in a double stranded state (page 19, Table A).”
(Id.). Abramson is relied on as teaching “the use of 5'-3' exonuclease deficient polymerases
in PCR (columns 2-8 and especially, column 7, lines 52-57).” (Answer, page 5).
The examiner concludes that (Answer, pages 5):
[i]t would have been prima facie obvious to one having ordinary skill in the art
at the time of the invention was made to combine the PCR amplification and
hybridization method of Link with the labels of either Parkhurst or Heller since
Parkhurst states "The double-labeled oligomer is very effective in signaling
hybridization (page 292, column 1, paragraph 2)." . . . It would further have been
obvious to combine the method of Link in view of either Parkhurst or Heller with
the use of exonuclease deficient polymerase as taught by Abramson since
Abramson states "When utilized in a PCR process with double-stranded primer
template complex, the 5' to 3' exonuclease activity of a DNA polymerase may
result in degradation of the 5'-end of the oligonucleotide primers. This activity
is not only undesirable in PCR, but also in second strand cDNA synthesis and
sequencing processes (column 7, lines 52-57)". Abramson solves the problem
of exonuclease activity by eliminating the exonuclease activity, as stated "Thus,
one aspect of this invention involves the generation of thermostable DNA
polymerase mutants displaying greatly reduced, attenuated or completely
eliminated 5' to 3' exonuclease activity.
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