Invention Grant
- Patent Title: Methods and reagents for combined PCR amplification
- Patent Title (中): 组合PCR扩增的方法和试剂
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Application No.: US12193667Application Date: 2008-08-18
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Publication No.: US07847076B2Publication Date: 2010-12-07
- Inventor: Paul E. Mayrand
- Applicant: Paul E. Mayrand
- Applicant Address: US CA Carlsbad
- Assignee: Life Technologies Corporation
- Current Assignee: Life Technologies Corporation
- Current Assignee Address: US CA Carlsbad
- Main IPC: C07H21/00
- IPC: C07H21/00 ; C07H21/02 ; C07H21/04 ; C12Q1/68 ; C12P19/34

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
- US20090258351A1 Methods and Reagents for Combined PCR Amplification Public/Granted day:2009-10-15
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