发明授权
- 专利标题: Hybridization and mismatch discrimination using oligonucleotides conjugated to minor groove binders
- 专利标题(中): 使用与小沟结合物缀合的寡核苷酸的杂交和错配鉴别
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申请号: US10127936申请日: 2002-04-22
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公开(公告)号: US06884584B2公开(公告)日: 2005-04-26
- 发明人: Joel Hedgpeth , Irina A. Afonina , Igor V. Kutyavin , Eugeny A. Lukhtanov , Evgeniy S. Belousov , Rich B. Meyer, Jr.
- 申请人: Joel Hedgpeth , Irina A. Afonina , Igor V. Kutyavin , Eugeny A. Lukhtanov , Evgeniy S. Belousov , Rich B. Meyer, Jr.
- 申请人地址: US WA Bothell
- 专利权人: Epoch Biosciences, Inc.
- 当前专利权人: Epoch Biosciences, Inc.
- 当前专利权人地址: US WA Bothell
- 代理机构: Townsend and Townsend and Crew LLP
- 主分类号: C12N15/09
- IPC分类号: C12N15/09 ; C07H19/23 ; C07H21/00 ; C07H21/02 ; C07H21/04 ; C12Q1/68 ; G01N21/78 ; G01N33/483 ; G01N33/53 ; G01N33/566 ; C07H19/04 ; C12P19/34
摘要:
Conjugates between a minor groove binding molecule, such as the trimer of 1,2-dihydro-(3H)-pyrrolo[3,2-e]indole-7-carboxylate (CDPI3), and an oligonucleotide form unusually stable hybrids with complementary target sequences, in which the tethered CDPI3 group resides in the minor groove of the duplex. These conjugates can be used as probes and primers. Due to their unusually high binding affinity, conjugates as short as 8-mers can be used as amplification primers with high specificity and efficiency. MGB conjugation also increases the discriminatory power of short oligonucleotides, providing enhanced detection of nucleotide sequence mismatches by short oligonucleotides. The MGB-conjugated probes and primers described herein facilitate various analytic and diagnostic procedures, such as amplification reactions, PCR, detection of single-nucleotide polymorphisms, gene hunting, differential display, fluorescence energy transfer, hydrolyzable probe assays and others; by allowing the use of shorter oligonucleotides, which have higher specificity and better discriminatory power.
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