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    Engineered templates and their use in single primer amplification
    3.
    发明授权
    Engineered templates and their use in single primer amplification 有权
    工程模板及其在单引物扩增中的应用

    公开(公告)号：US07306906B2

    公开(公告)日：2007-12-11

    申请号：US10251085

    申请日：2002-09-19

    申请人： Toshiaki Maruyama ,  Shana Frederickson ,  Katherine S. Bowdish ,  Mark Renshaw ,  Ying-Chi Lin

    发明人： Toshiaki Maruyama ,  Shana Frederickson ,  Katherine S. Bowdish ,  Mark Renshaw ,  Ying-Chi Lin

    IPC分类号： C12Q1/68 ,  C12P19/34 ,  C07H21/02 ,  C07H21/04

    CPC分类号： C12N15/1093 ,  C12Q1/6844 ,  C12Q1/6853 ,  C12Q1/6855 ,  C12Q2537/157 ,  C12Q2533/101 ,  C12Q2525/301 ,  C12Q2525/186 ,  C12Q2525/161 ,  C12Q2525/155 ,  C12Q2525/131

    摘要： Methods of amplifying nucleic acid have now been discovered which include the steps of: a) annealing a primer to a template nucleic acid sequence, the primer having a first portion which anneals to the template and a second portion of predetermined sequence; b) synthesizing a polynucleotide that anneals to and is complementary to the portion of the template between the location at which the first portion of the primer anneals to the template and the end of the template, the polynucleotide having a first end and a second end, wherein the first end incorporates the primer; c) separating the polynucleotide synthesized in step (b) from the template; d) annealing a nested oligonucleotide to the second end of the polynucleotide synthesized in step (b), the nested oligonucleotide having a first portion that anneals to the second end of the polynucleotide and a second portion having the same predetermined sequence as the second portion of the primer; e) extending the polynucleotide synthesized in step (b) to provide a terminal portion thereof that is complementary to the predetermined sequence; and f) amplifying the extended polynucleotide using a single primer having the predetermined sequence.

    摘要翻译： 现已发现扩增核酸的方法，其包括以下步骤：a）将引物退火至模板核酸序列，所述引物具有与模板退火的第一部分和预定序列的第二部分; b）合成在所述模板的第一部分与所述模板退火的位置与所述模板的末端之间退火并与所述模板部分互补的多核苷酸，所述多核苷酸具有第一末端和第二末端， 其中所述第一端包含所述底漆; c）将步骤（b）中合成的多核苷酸与模板分离; d）将嵌套的寡核苷酸退火到步骤（b）中合成的多核苷酸的第二末端，所述嵌套的寡核苷酸具有与所述多核苷酸的第二末端退火的第一部分和具有与所述多核苷酸的第二部分相同的预定序列的第二部分 底漆; e）延伸在步骤（b）中合成的多核苷酸以提供与预定序列互补的其末端部分; 和f）使用具有预定序列的单一引物扩增延伸的多核苷酸。

    NESTED OLIGONUCLEOTIDES CONTAINING A HAIRPIN FOR NUCLEIC ACID AMPLIFICATION
    4.
    发明申请
    NESTED OLIGONUCLEOTIDES CONTAINING A HAIRPIN FOR NUCLEIC ACID AMPLIFICATION 失效
    含有乳酸的核苷酸寡核苷酸用于核酸放大

    公开(公告)号：US20050079489A1

    公开(公告)日：2005-04-14

    申请号：US10014012

    申请日：2001-12-10

    申请人： Katherine Bowdish ,  Shana Frederickson ,  John McWhirter ,  Maruyama Toshiaki

    发明人： Katherine Bowdish ,  Shana Frederickson ,  John McWhirter ,  Maruyama Toshiaki

    IPC分类号： C12Q1/68 ,  C12P19/34

    CPC分类号： C12Q1/6844 ,  C12Q2525/301

    摘要： Templates that are engineered to contain a predetermined sequence and a hairpin structure are provided by a nested oligonucleotide extension reaction. The engineered template allows Single Primer Amplification (SPA) to amplify a target sequence within the engineered template. In particularly useful embodiments, the target sequences from the engineered templates are cloned into expression vehicles to provide a library of polypeptides or proteins, such as, for example, an antibody library.

    摘要翻译： 通过嵌套的寡核苷酸延伸反应提供被设计成含有预定序列和发夹结构的模板。 工程模板允许单引物扩增（SPA）扩增工程模板内的靶序列。 在特别有用的实施方案中，将来自工程化模板的靶序列克隆到表达载体中以提供多肽或蛋白质文库，例如抗体文库。

    Engineered templates and their use in single primer amplification
    7.
    发明授权
    Engineered templates and their use in single primer amplification 有权
    工程模板及其在单引物扩增中的应用

    公开(公告)号：US07414111B2

    公开(公告)日：2008-08-19

    申请号：US10737252

    申请日：2003-12-15

    申请人： Toshiaki Maruyama ,  Katherine S. Bowdish ,  Shana Frederickson ,  Mark Renshaw

    发明人： Toshiaki Maruyama ,  Katherine S. Bowdish ,  Shana Frederickson ,  Mark Renshaw

    IPC分类号： C07K16/00

    CPC分类号： C07K16/4291 ,  C07K16/005 ,  C07K16/082 ,  C07K2317/55 ,  C07K2317/56 ,  C07K2317/565 ,  C12N15/1093 ,  C12Q1/6844 ,  C12Q1/6853 ,  C12Q1/6855 ,  C12Q1/686 ,  Y10S530/861 ,  C12Q2525/161 ,  C12Q2525/155 ,  C12Q2537/157 ,  C12Q2533/101 ,  C12Q2525/301 ,  C12Q2525/131 ,  C12Q2525/186

    摘要： Methods of amplifying nucleic acid have now been discovered which include the steps of: a) annealing a primer to a template nucleic acid sequence, the primer having a first portion which anneals to the template and a second portion of predetermined sequence; b) synthesizing a polynucleotide that anneals to and is complementary to the portion of the template between the location at which the first portion of the primer anneals to the template and the end of the template, the polynucleotide having a first end and a second end, wherein the first end incorporates the primer; c) separating the polynucleotide synthesized in step (b) from the template; d) annealing a nested oligonucleotide to the second end of the polynucleotide synthesized in step (b), the nested oligonucleotide having a first portion that anneals to the second end of the polynucleotide and a second portion having the same predetermined sequence as the second portion of the primer; e) extending the polynucleotide synthesized in step (b) to provide a terminal portion thereof that is complementary to the predetermined sequence; and f) amplifying the extended polynucleotide using a single primer having the predetermined sequence. In particularly useful embodiments, the methods are used to amplify a repertoire of IgA antibodies.

    摘要翻译： 现已发现扩增核酸的方法，其包括以下步骤：a）将引物退火至模板核酸序列，所述引物具有与模板退火的第一部分和预定序列的第二部分; b）合成在所述模板的第一部分与所述模板退火的位置与所述模板的末端之间退火并与所述模板部分互补的多核苷酸，所述多核苷酸具有第一末端和第二末端， 其中所述第一端包含所述底漆; c）将步骤（b）中合成的多核苷酸与模板分离; d）将嵌套的寡核苷酸退火到步骤（b）中合成的多核苷酸的第二末端，所述嵌套的寡核苷酸具有与所述多核苷酸的第二末端退火的第一部分和具有与所述多核苷酸的第二部分相同的预定序列的第二部分 底漆; e）延伸在步骤（b）中合成的多核苷酸以提供与预定序列互补的其末端部分; 和f）使用具有预定序列的单一引物扩增延伸的多核苷酸。 在特别有用的实施方案中，所述方法用于扩增IgA抗体的所有组成成分。