发明授权
US07014994B1 Coupled polymerase chain reaction-restriction-endonuclease digestion-ligase detection reaction process
有权
耦合聚合酶链反应 - 限制性内切酶消化 - 连接酶检测反应过程
- 专利标题: Coupled polymerase chain reaction-restriction-endonuclease digestion-ligase detection reaction process
- 专利标题(中): 耦合聚合酶链反应 - 限制性内切酶消化 - 连接酶检测反应过程
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申请号: US09528014申请日: 2000-03-17
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公开(公告)号: US07014994B1公开(公告)日: 2006-03-21
- 发明人: Francis Barany , Joseph P. Day , Robert P. Hammer , Donald E. Bergstrom
- 申请人: Francis Barany , Joseph P. Day , Robert P. Hammer , Donald E. Bergstrom
- 申请人地址: US NY Ithaca US IN West Lafayette US LA Baton Rouge
- 专利权人: Cornell Research Foundation,Inc.,Purdue Research Foundation,Board of Supervisors of Louisiana State University and Agricultural and Mechanical College
- 当前专利权人: Cornell Research Foundation,Inc.,Purdue Research Foundation,Board of Supervisors of Louisiana State University and Agricultural and Mechanical College
- 当前专利权人地址: US NY Ithaca US IN West Lafayette US LA Baton Rouge
- 代理机构: Nixon Peabody LLP
- 主分类号: C12Q1/68
- IPC分类号: C12Q1/68 ; C12P19/34 ; C07H21/04
摘要:
The present invention provides a method for identifying one or more low abundance sequences differing by one or more single-base changes, insertions, or deletions, from a high abundance sequence in a plurality of target nucleotide sequences. The high abundance wild-type sequence is selectively removed using high fidelity polymerase chain reaction analog conversion, facilitated by optimal buffer conditions, to create a restriction endonuclease site in the high abundance wild-type gene, but not in the low abundance mutant gene. This allows for digestion of the high abundance DNA. Subsequently the low abundant mutant DNA is amplified and detected by the ligase detection reaction assay. The present invention also relates to a kit for carrying out this procedure.
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