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专利名称：Methods and reagents for combined PCR
amplification and hybridization probing
发明人：Paul E. Mayrand申请号：US10293236申请日：20021112
公开号：US20030215826A1公开日：20031120
摘要：An oligonucleotide probe is disclosed, the probe including an oligonucleotide, afluorescer molecule attached to a first end of the oligonucleotide and a quenchermolecule attached to the opposite end of the oligonucleotide. The probe is renderedimpervious to digestion by the 5′→3′ exonuclease activity of a polymerase and the 3′→5′extension of by a polymerase. The invention also includes methods for performingcombined PCR amplification and hybridization probing, one such method including thesteps of contacting a target nucleic acid sequence with PCR reagents and an
oligonucleotide probe as described above, and subjecting these reagents to thermalcycling. One preferred refinement of the above method further includes the addition of astrand displacer to facilitate amplification. Additional similar combined PCR hybridizationmethods are disclosed, such methods not requiring probes having their 5′ endsprotected, wherein (i) the polymerase lacks 5′→3′ exonuclease activity, (ii) a 5′→3′exonuclease inhibitor is included, and (iii) an exonuclease deactivation step is performed.
申请人：PE CORPORATION (NY)
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