C - Chemistry – Metallurgy – 12 – Q
Patent
C - Chemistry, Metallurgy
12
Q
C12Q 1/68 (2006.01) C12N 15/09 (2006.01) C12P 19/34 (2006.01) G01N 33/58 (2006.01)
Patent
CA 2472970
The present invention is directed to a method and a composition for amplifying and detecting a target nucleic comprising a) subjecting said target nucleic acid to a real time PCR amplification reaction in the presence of (i) a thermostable DNA Polymerase, (ii) a thermostable double strand dependent 3'-5' exonuclease having a temperature optimum above 37°C , (iii) a pair of amplification primers, (iv) Deoxynucleoside-Tri-Phosphates, (v) a detecting oligonucleotide carrying a first label and a second label, said first label being capable of acting as a fluorescent reporter entity when excited with light of an appropriate wavelength, said second label being capable of acting as a fluorescence quenching entity of said fluorescent reporter entity, characterized in that one label is bound to the 3' end of said detecting oligonucleotide, and further characterized in that the other label is bound either internally or at the 5' end to said detecting oligonucleotide, and b) monitoring fluorescence of said fluorerescent reporter entity at least after a plurality of amplification cycles.
Ankenbauer Waltraud
Heindl Dieter
Laue Frank
F.hoffmann-La Roche Ag
Norton Rose Or S.e.n.c.r.l. S.r.l./llp
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