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    Method for amplifying target nucleic acid sequences using a primer comprising an AP endonuclease-cleavable linker
    4.
    发明授权
    Method for amplifying target nucleic acid sequences using a primer comprising an AP endonuclease-cleavable linker 有权
    使用包含AP内切核酸酶切割接头的引物扩增靶核酸序列的方法

    公开(公告)号：US07553643B2

    公开(公告)日：2009-06-30

    申请号：US11432763

    申请日：2006-05-10

    申请人： Igor V. Kutyavin ,  David Milesi ,  Merl F. Hoekstra

    发明人： Igor V. Kutyavin ,  David Milesi ,  Merl F. Hoekstra

    IPC分类号： C12P19/34 ,  C12Q1/68 ,  C07H21/00

    CPC分类号： C12Q1/6813 ,  C12Q1/6827 ,  C12Q2525/119 ,  C12Q2521/313

    摘要： The present invention provides a novel method for nucleic acid amplification. The method includes contacting the sample with at least one forward primer and at least one reverse primer, an AP endonuclease, and a nucleic acid polymerase. under conditions sufficient to allow the forward and reverse primers to hybridize to the target nucleic acid and form a reaction mixture, wherein at least one of the forward and reverse primer includes an AP endonuclease-cleavable linker L, and incubating the reaction mixture under reaction conditions that simultaneously allow the AP endonuclease to cleave at a linker site L and the extension of the primers in a template-specific manner to amplify the target nucleic acid sequence.

    摘要翻译： 本发明提供了一种新的核酸扩增方法。 该方法包括使样品与至少一个正向引物和至少一个反向引物，AP内切核酸酶和核酸聚合酶接触。 在足以允许正向和反向引物与靶核酸杂交并形成反应混合物的条件下，其中正向引物和反向引物中的至少一个引物包含AP内切核酸酶可切割连接体L，并在反应条件下温育反应混合物 其同时允许AP内切核酸酶在连接位点L处切割，并以模板特异性方式扩增引物以扩增靶核酸序列。

    OLIGONUCLEOTIDES CONTAINING PYRAZOLO[3,4-D] PYRIMIDINES FOR HYBRIDIZATION AND MISMATCH DISCRIMINATION
    5.
    发明申请
    OLIGONUCLEOTIDES CONTAINING PYRAZOLO[3,4-D] PYRIMIDINES FOR HYBRIDIZATION AND MISMATCH DISCRIMINATION 审中-公开
    含有吡唑并[3,4-D]嘧啶的混合物用于杂交和错配歧视的寡核苷酸

    公开(公告)号：US20100228017A1

    公开(公告)日：2010-09-09

    申请号：US11970445

    申请日：2008-01-07

    申请人： Rich B. Meyer, JR. ,  Irina A. Afonina ,  Igor V. Kutyavin

    发明人： Rich B. Meyer, JR. ,  Irina A. Afonina ,  Igor V. Kutyavin

    IPC分类号： C07H21/00

    CPC分类号： C12Q1/6818 ,  C12Q1/6827 ,  C12Q1/6832 ,  C12Q2527/107 ,  C12Q2525/117

    摘要： Oligonucleotides in which one or more purine residues are substituted by pyrazolo[3,4-d]pyrimidines exhibit improved hybridization properties. Oligonucleotides containing pyrazolo[3,4-d]pyrimidine base analogues have higher melting temperatures than unsubstituted oligonucleotides of identical sequence. Thus, in assays involving hybridization of an oligonucleotide probe to a target polynucleotide sequence, higher signals are obtained. In addition, mismatch discrimination is enhanced when pyrazolo[3,4-d]pyrimidine-containing oligonucleotides are used as hybridization probes, making them useful as probes and primers for hybridization, amplification and sequencing procedures, particularly those in which single- or multiple-nucleotide mismatch discrimination is required.

    摘要翻译： 其中一个或多个嘌呤残基被吡唑并[3,4-d]嘧啶取代的寡核苷酸表现出改善的杂交性质。 含有吡唑并[3,4-d]嘧啶碱基类似物的寡核苷酸具有比相同序列的未取代寡核苷酸更高的融合温度。 因此，在涉及寡核苷酸探针杂交到靶多核苷酸序列的测定中，获得较高的信号。 此外，当使用含有吡唑并[3,4-d]嘧啶的寡核苷酸作为杂交探针时，错配鉴别被增强，使其可用作用于杂交，扩增和测序程序的探针和引物，特别是单链或多 需要核苷酸错配鉴别。

    Oligonucleotides including pyrazolo[3,4-D]pyrimidine bases, bound in double stranded nucleic acids
    8.
    发明授权
    Oligonucleotides including pyrazolo[3,4-D]pyrimidine bases, bound in double stranded nucleic acids 失效
    寡核苷酸包括双链核酸结合的吡唑并[3,4-D]嘧啶碱基

    公开(公告)号：US6143877A

    公开(公告)日：2000-11-07

    申请号：US848373

    申请日：1997-04-30

    申请人： Rich B. Meyer ,  Alexander Gall ,  Igor V. Kutyavin

    发明人： Rich B. Meyer ,  Alexander Gall ,  Igor V. Kutyavin

    IPC分类号： C07H21/00 ,  C12Q1/68 ,  C07H21/02 ,  C07H21/04

    CPC分类号： C07H21/00 ,  C12Q1/6813

    摘要： A triplex forming oligonucleotide is complementary pursuant to the G/T or A/G recognition motif to a homopurine, or substantially homopurine target sequence in double stranded nucleic acids, and at least one and preferably all of the guanine bases are replaced by their pyrazolo[3,4-d]pyrimidine analog, namely by 6-amino-1H-pyrazolo[3,4-d]pyrimidin-4(5H)-one. The oliginucleotides containing the pyrazolo[3,4-d]pyrimidine analog of guanine exhibit a lesser degree of self-association, and lack the nucleophilic nitrogen atom in the 7 position of guanine. The latter feature results in a diminished extent of self-crosslinking in ODNs which also have a covalently attached cross-linking agent.

    摘要翻译： 三链体形成寡核苷酸是根据G / T或A / G识别基序与双链核酸中的均嘌呤或基本上均嘌呤靶序列互补的，并且至少一个并优选全部鸟嘌呤碱基被其吡唑并[ 3,4-d]嘧啶类似物，即6-氨基-1H-吡唑并[3,4-d]嘧啶-4（5H） - 酮。 含有鸟嘌呤的吡唑并[3,4-d]嘧啶类似物的寡核苷酸表现出较小程度的自身结合，并且在鸟嘌呤的7位缺乏亲核氮原子。 后一个特征导致ODN中的自交联程度减小，其也具有共价连接的交联剂。

    Hybridization and mismatch discrimination using oligonucleotides conjugated to minor groove binders
    10.
    发明授权
    Hybridization and mismatch discrimination using oligonucleotides conjugated to minor groove binders 有权
    使用与小沟结合物缀合的寡核苷酸的杂交和错配鉴别

    公开(公告)号：US07794945B2

    公开(公告)日：2010-09-14

    申请号：US12172999

    申请日：2008-07-14

    申请人： Joel Hedgpeth ,  Irina A. Afonina ,  Igor V. Kutyavin ,  Eugeny A. Lukhtanov ,  Evgeniy S. Belousov ,  Rich B. Meyer, Jr.

    发明人： Joel Hedgpeth ,  Irina A. Afonina ,  Igor V. Kutyavin ,  Eugeny A. Lukhtanov ,  Evgeniy S. Belousov ,  Rich B. Meyer, Jr.

    IPC分类号： C12Q1/68 ,  C07H21/02 ,  C07H21/04

    CPC分类号： C07H21/00 ,  C07D207/34 ,  C07D403/12 ,  C07D487/04 ,  C07H19/073 ,  C07H19/173 ,  C07H19/23 ,  C07H21/04 ,  C12Q1/6813 ,  C12Q2527/107 ,  C12Q2525/197

    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.

    摘要翻译： 小沟结合分子（例如1,2-二氢 - （3H） - 吡咯并[3,2-e]吲哚-7-羧酸酯（CDPI3）的三聚体和寡核苷酸之间的缀合物形成非常稳定的具有互补靶的杂交体 序列，其中系留的CDPI3基团位于双链体的小沟槽中。 这些缀合物可以用作探针和引物。 由于其非常高的结合亲和力，短至8聚体的缀合物可以用作高特异性和高效的扩增引物。 MGB缀合还增加了短寡核苷酸的鉴别能力，提供了通过短寡核苷酸增强核苷酸序列错配检测。 本文所述的MGB缀合的探针和引物有助于各种分析和诊断程序，如扩增反应，PCR，单核苷酸多态性检测，基因敲除，差异显示，荧光能量转移，水解性探针测定等。 通过允许使用具有更高特异性和更好的辨别力的较短的寡核苷酸。