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    ENGINEERED TEMPLATES AND THEIR USE IN SINGLE PRIMER AMPLIFICATION
    5.
    发明申请
    ENGINEERED TEMPLATES AND THEIR USE IN SINGLE PRIMER AMPLIFICATION 审中-公开
    工程模板及其在单一放大器中的使用

    公开(公告)号：WO03025202A2

    公开(公告)日：2003-03-27

    申请号：PCT/US0229889

    申请日：2002-09-19

    Applicant: ALEXION PHARMA INC ,  BOWDISH KATHERINE S ,  FREDERICKSON SHANA ,  MARUYAMA TOSHIAKI ,  LIN YING-CHI ,  RENSHAW MARK

    Inventor： BOWDISH KATHERINE S ,  FREDERICKSON SHANA ,  MARUYAMA TOSHIAKI ,  LIN YING-CHI ,  RENSHAW MARK

    IPC: C12N15/09 ,  C12N15/10 ,  C12Q1/68 ,  C12Q

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

    Abstract: 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.

    Abstract translation: 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。

    ENGINEERED TEMPLATES AND THEIR USE IN SINGLE PRIMER AMPLIFICATION
    7.
    发明申请
    ENGINEERED TEMPLATES AND THEIR USE IN SINGLE PRIMER AMPLIFICATION 审中-公开
    工程模板及其在单一放大器中的使用

    公开(公告)号：WO2005060641A3

    公开(公告)日：2008-02-21

    申请号：PCT/US2004041945

    申请日：2004-12-15

    Applicant: ALEXION PHARMA INC ,  MARUYAMA TOSHIAKI ,  FREDERICKSON SHANA ,  BOWDISH KATHERINE S ,  RENSHAW MARK ,  LIN YING-CHI

    Inventor： MARUYAMA TOSHIAKI ,  FREDERICKSON SHANA ,  BOWDISH KATHERINE S ,  RENSHAW MARK ,  LIN YING-CHI

    IPC: C12Q1/68 ,  C07K16/00 ,  C07K16/08 ,  C07K16/42 ,  C12N15/10 ,  C12N15/11 ,  C12P19/34

    CPC classification number: 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

    Abstract: 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.

    Abstract translation: 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。

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