Invention Patent
- Patent Title: Methods and reagents therefor which combines Pcr amplification and hybridization probing assays
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Application No.: JP2003099758Application Date: 2003-04-02
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Publication No.: JP4040519B2Publication Date: 2008-01-30
- Inventor: イー. メイランド ポール
- Applicant: アプレラ コーポレイション
- Assignee: アプレラ コーポレイション
- Current Assignee: アプレラ コーポレイション
- Priority: US43550995 1995-05-05
- Main IPC: C12N15/09
- IPC: C12N15/09 ; C07H21/04 ; C12P19/34 ; C12Q1/68 ; G01N21/64 ; G01N21/77 ; G01N21/78 ; G01N33/53 ; G01N33/566
Abstract:
An oligonucleotide probe is disclosed, the probe including an oligonucleotide, a fluorescer molecule attached to a first end of the oligonucleotide and a quencher molecule attached to the opposite end of the oligonucleotide. The probe is rendered impervious to digestion by the 5′→3′ exonuclease activity of a polymerase and the 5′→3′ extension of by a polymerase. The invention also includes methods for performing combined PCR amplification and hybridization probing, one such method including the steps of contacting a target nucleic acid sequence with PCR reagents and an oligonucleotide probe as described above, and subjecting these reagents to thermal cycling. One preferred refinement of the above method further includes the addition of a strand displacer to facilitate amplification. Additional similar combined PCR hybridization methods are disclosed, such methods not requiring probes having their 5′ ends protected, 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.
Public/Granted literature
- JP2004000185A Method for combining pcr amplification with hybridization probing assay and reagent threfor Public/Granted day:2004-01-08
Information query
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