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    Detection of single nucleotide polymorphisms
    1.
    发明专利
    Detection of single nucleotide polymorphisms 未知

    公开(公告)号：AU3803001A

    公开(公告)日：2001-08-14

    申请号：AU3803001

    申请日：2001-02-02

    Applicant: ADVION BIOSCIENCES INC

    Inventor： SCHULTZ GARY A ,  ZHANG SHENG ,  PELT COLLEEN K VAN

    IPC: C12Q1/68

    Abstract: The present invention relates to a method of detecting single nucleotide polymorphisms by providing a target nucleic acid molecule, an oligonucleotide primer complementary to a portion of the target nucleic acid molecule, a nucleic acid polymerizing enzyme, and a plurality of types of nucleotide analogs. The target nucleic molecule, the oligonucleotide primer, the nucleic acid polymerizing enzyme, and the nucleotide analogs, each type being present in a first amount, are blended to form an extension solution where the oligonucleotide primer is hybridized to the target nucleic acid molecule to form a primed target nucleic acid molecule and the nucleic acid polymerizing enzyme is positioned to add nucleotide analogs to the primed target nucleic acid molecule at an active site. The oligonucleotide primer in the extension solution is extended by using the nucleic acid polymerizing enzyme to add a nucleotide analog to the oligonucleotide primer at the active site. This forms an extended oligonucleotide primer, wherein the nucleotide analog being added is complementary to the nucleotide of the target nucleic acid molecule at the active site. The amounts of each type of the nucleotide analogs in the extension solution after the extending step are then determined where each type is present in a second amount. The first and second amounts of each type of the nucleotide analog are compared. The type of nucleotide analog where the first and second amounts differ as the nucleotide added to the oligonucleotide primer at the active site is then identified. The steps of extending, determining the amounts of each type of the nucleotide analog, comparing the first and second amounts of the nucleotide analog, and said identifying the type of nucleotide analog added may be repeated, either after repeating the blending with the extended oligonucleotide primer or after determining the amounts of each type of dideoxynucleotide or dideoxynucleotide analog remaining in the extension solution as the new first amount. As a result, the nucleotide at the active site of the target nucleic acid molecule is determined. Also disclosed is an apparatus and composition for carrying out this method.

    A ONE-WELL ASSAY FOR HIGH THROUGHPUT DETECTION OF SINGLE NUCLEOTIDE POLYMORPHISMS
    4.
    发明申请
    A ONE-WELL ASSAY FOR HIGH THROUGHPUT DETECTION OF SINGLE NUCLEOTIDE POLYMORPHISMS 审中-公开
    高通量检测单核苷酸多态性的一种检测方法

    公开(公告)号：WO0234883A3

    公开(公告)日：2002-11-07

    申请号：PCT/US0150857

    申请日：2001-10-27

    Applicant: ADVION BIOSCIENCES INC

    Inventor： ZHANG SHENG ,  VAN PELT COLLEEN K ,  SCHULTZ GARY A

    IPC: C12Q1/68

    CPC classification number: C12Q1/6827 ,  C12Q1/6858 ,  C12Q2535/125 ,  C12Q2531/113 ,  C12Q2521/525 ,  C12Q2565/501

    Abstract: The present invention relates to a novel one-well assay which couples a conventional polymerase chain reaction (PCR) amplification step to a single nucleotide primer extension step for the the determination of nucleotide sequence variations in the genotyping of single nucleotide polymorphisms and other DNA variations detected by primer extension methods. A PCR amplification step, a phosphatase digestion step (or a molecular weight-selective filter step), and a primer extension step are consecutively performed in the same well plate followed by electrospray mass spectrometry detection of a single nucleotide polymorphism bases. Alternative one-well assays which utilize exonuclease I or lambda -exonuclease in addition to the phosphatase digestion step are also disclosed.

    Abstract translation: 本发明涉及将常规聚合酶链式反应（PCR）扩增步骤偶联至单核苷酸引物延伸步骤以用于确定单核苷酸多态性和检测到的其他DNA变异的基因型分析中的核苷酸序列变异的新颖单孔试验 通过引物延伸方法。 在同一孔板中连续进行PCR扩增步骤，磷酸酶消化步骤（或分子量选择性过滤步骤）和引物延伸步骤，接着通过电喷雾质谱检测单核苷酸多态性碱基。 也公开了除了磷酸酶消化步骤之外还使用外切核酸酶I或λ-外切核酸酶的替代性单孔测定法。

    DETECTION OF SINGLE NUCLEOTIDE POLYMORPHISMS
    6.
    发明公开
    DETECTION OF SINGLE NUCLEOTIDE POLYMORPHISMS 审中-公开
    单检测

    公开(公告)号：EP1252336A4

    公开(公告)日：2005-02-09

    申请号：EP01910423

    申请日：2001-02-02

    Applicant: ADVION BIOSCIENCES INC

    Inventor： SCHULTZ GARY A ,  ZHANG SHENG ,  VAN PELT COLLEEN K

    IPC: C12Q1/68

    CPC classification number: C12Q1/6858 ,  B01J2219/00274 ,  C12Q1/6869 ,  H01J49/165 ,  C12Q2565/627

    Abstract: The present invention relates to a method of detecting single nucleotide polymorphisms by providing a target nucleic acid molecule, an oligonucleotide primer complementary to a portion of the target nucleic acid molecule, a nucleic acid polymerizing enzyme, and a plurality of types of nucleotide analogs. The target nucleic molecule, the oligonucleotide primer, the nucleic acid polymerizing enzyme, and the nucleotide analogs, each type being present in a first amount, are blended to form an extension solution where the oligonucleotide primer is hybridized to the target nucleic acid molecule to form a primed target nucleic acid molecules and the nucleic acid polymerizing enzyme is positioned to add nucleotide analogs to the prime target nucleic acid molecule at an active site. The oligonucleotide primer in the extension solution is extended by using the nucleic acid polymerizing enzyme to add a nucleotide analog to the oligonucleotide primer at the active site. This forms an extended oligonucleotide primer, wherein the nucleotide analog being added is complementary to the nucleotide of the target nucleic acid molecule at the active site. The amounts of each type of the nucleotide analogs in the extension solution after the extending step are then determined where each type is present in a second amount. The first and second amounts of each type of the nucleotide analog are compared. The type of nucleotide analog where the first and second amounts differ as the nucleotide added to the oligonucleotide primer at the active site is then identified. The steps of extending, determining the amounts of each type of the nucleotide analog, comparing the first and second amounts of the nucleotide analog, and said identifying the type of nucleotide analog added may be repeated, either after repeating the blending with the extended oligonucleotide primer or after determining the amounts of each type of dideoxynucleotide or dideoxynucleotide analog remaining in the extension solution as the new first amount. As a result, the nucleotide at the active site of the target nucleic acid molecule is determined. Also disclosed is an apparatus and composition for carrying out this method.

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