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    Optimal procedure for isolation of mutant mitochondrial alleles
    2.
    发明授权
    Optimal procedure for isolation of mutant mitochondrial alleles 失效
    分离突变体线粒体等位基因的最佳方法

    公开(公告)号：US6027883A

    公开(公告)日：2000-02-22

    申请号：US757438

    申请日：1996-11-27

    Applicant: Corinna Herrnstadt ,  Soumitra Ghosh ,  Eoin D. Fahy ,  Robert E. Davis

    Inventor： Corinna Herrnstadt ,  Soumitra Ghosh ,  Eoin D. Fahy ,  Robert E. Davis

    IPC: C12N15/09 ,  C12Q1/68

    CPC classification number: C12Q1/6883 ,  C12Q1/6806 ,  C12Q1/6858 ,  C12Q2600/156 ,  C12Q2600/16

    Abstract: A sample preparation procedure for mitochondrial (mt) DNA analysis is described. The present method for isolating mtDNA uses sedimentation techniques for separating erythrocytes from lymphocytes and platelets (crude buffy coat fraction), followed by DNA extraction from the crude buffy coat fraction by boiling in water. This approach improves the yield of mutant DNA, enhancing the sensitivity of subsequent mutation interrogation techniques and allowing for meaningful statistical treatment of the degree of heteroplasmy within the mitochondrial DNA.

    Abstract translation: 描述了线粒体（mt）DNA分析的样品制备程序。 目前用于分离mtDNA的方法使用沉淀技术从淋巴细胞和血小板（粗血沉棕黄层级分离）中分离红细胞，然后通过在水中沸腾从粗血沉棕黄层级分提取DNA。 这种方法提高了突变DNA的产量，提高了随后的突变询问技术的敏感性，并允许有意义的统计学处理线粒体DNA内的异质性程度。

    End-attachment of oligonucleotides to polyacrylamide solid supports for capture and detection of nucleic acids
    3.
    发明授权
    End-attachment of oligonucleotides to polyacrylamide solid supports for capture and detection of nucleic acids 失效
    将寡核苷酸末端连接到用于捕获和检测核酸的聚丙烯酰胺固体支持物上

    公开(公告)号：US5663242A

    公开(公告)日：1997-09-02

    申请号：US414019

    申请日：1995-03-31

    Applicant: Soumitra Shanker Ghosh ,  Eoin D. Fahy

    Inventor： Soumitra Shanker Ghosh ,  Eoin D. Fahy

    IPC: C07H21/00 ,  C12Q1/68 ,  C08F20/56 ,  C12P19/34 ,  C12Q1/70

    CPC classification number: C07H21/00

    Abstract: Methods for covalent attachment of oligonucleotides to solid supports such that substantially all of the oligonucleotides are attached via their 5'-ends are provided. The solid supports with attached oligonucleotides are produced. Thiol-oligonucleotides are attached to bromoacetyl-derivatized polyacrylamide supports, or conversely, bromoacetyl-oligonucleotides are immobilized on thiol-polyacrylamide supports. In a further aspect, bromoacetyl-derivatized oligonucleotides, and polyacrylamide supports with linked oligonucleotides produced by coupling bromoacetyl-derivatized oligonucleotides with thiol-derivatized polyacrylamide solid supports or by coupling thiol-derivatized oligonucleotides with bromoacetyl-derivatized polyacrylamide supports as well as methods for capture of nucleic acids by oligonucleotides attached to polyacrylamide solid supports, either by direct capture or in sandwich hybridization formats are provided.

    Abstract translation: 提供了将寡核苷酸共价连接到固体支持物的方法，使得基本上所有的寡核苷酸通过其5'-末端连接。 产生具有连接的寡核苷酸的固体支持物。 硫醇寡核苷酸连接到溴乙酰基衍生的聚丙烯酰胺载体上，或者相反地，溴乙酰基寡核苷酸固定在硫醇 - 聚丙烯酰胺载体上。 在另一方面，溴乙酰基衍生的寡核苷酸和具有通过将溴乙酰基衍生的寡核苷酸与硫醇衍生的聚丙烯酰胺固体支持物偶联或通过将硫醇衍生的寡核苷酸与溴乙酰基衍生的聚丙烯酰胺载体偶联产生的链接寡核苷酸的聚丙烯酰胺载体以及捕获 提供了通过直接捕获或以夹心杂交形式连接到聚丙烯酰胺固体支持物上的寡核苷酸的核酸。

    End-attachment of oligonucleotides to polyacrylamide solid supports for capture and detection of nucleic acids
    4.
    发明授权
    End-attachment of oligonucleotides to polyacrylamide solid supports for capture and detection of nucleic acids 失效
    将寡核苷酸末端连接到用于捕获和检测核酸的聚丙烯酰胺固体支持物上

    公开(公告)号：US5478893A

    公开(公告)日：1995-12-26

    申请号：US102474

    申请日：1993-08-05

    Applicant: Soumitra S. Ghosh ,  Eoin D. Fahy

    Inventor： Soumitra S. Ghosh ,  Eoin D. Fahy

    IPC: C07H21/00 ,  C12Q1/68 ,  C08F120/56 ,  C07H21/02 ,  C07H21/04

    CPC classification number: C07H21/00

    Abstract: Methods for covalent attachment of oligonucleotides to solid supports such that substantially all of the oligonucleotides are attached via their 5'-ends are provided. The solid supports with attached oligonucleotides are produced. Thiol-oligonucleotides are attached to bromoacetyl-derivatized polyacrylamide supports, or conversely, bromoacetyl-oligonucleotides are immobilized on thiol-polyacrylamide supports. In a further aspect, bromoacetyl-derivatized oligonucleotides, and polyacrylamide supports with linked oligonucleotides produced by coupling bromoacetyl-derivatized oligonucleotides with thiol-derivatized polyacrylamide solid supports or by coupling thiol-derivatized oligonucleotides with bromoacetyl-derivatized polyacrylamide supports as well as methods for capture of nucleic acids by oligonucleotides attached to polyacrylamide solid supports, either by direct capture or in sandwich hybridization formats are provided.

    Abstract translation: 提供了将寡核苷酸共价连接到固体支持物的方法，使得基本上所有的寡核苷酸通过其5'-末端连接。 产生具有连接的寡核苷酸的固体支持物。 硫醇寡核苷酸连接到溴乙酰基衍生的聚丙烯酰胺载体上，或者相反地，溴乙酰基寡核苷酸固定在硫醇 - 聚丙烯酰胺载体上。 在另一方面，溴乙酰基衍生的寡核苷酸和具有通过将溴乙酰基衍生的寡核苷酸与硫醇衍生的聚丙烯酰胺固体支持物偶联或通过将硫醇衍生的寡核苷酸与溴乙酰基衍生的聚丙烯酰胺载体偶联产生的链接寡核苷酸的聚丙烯酰胺载体以及捕获 提供了通过直接捕获或以夹心杂交形式连接到聚丙烯酰胺固体支持物上的寡核苷酸的核酸。

    End-attachment of oligonucleotides to polyacrylamide solid supports for capture and detection of nucleic acids
    7.
    发明授权
    End-attachment of oligonucleotides to polyacrylamide solid supports for capture and detection of nucleic acids 失效
    将寡核苷酸结合到聚丙烯酰胺固体支持物上用于捕获和检测核酸

    公开(公告)号：US5237016A

    公开(公告)日：1993-08-17

    申请号：US293893

    申请日：1989-01-05

    Applicant: Soumitra S. Ghosh ,  Eoin D. Fahy

    Inventor： Soumitra S. Ghosh ,  Eoin D. Fahy

    IPC: C07H21/00 ,  C12Q1/68

    CPC classification number: C07H21/00

    Abstract: This invention concerns methods and means for covalent attachment of oligonucleotides to solid supports substantially at their 5'-ends. According to the invention thiol-oligonucleotides are attached to bromoacetyl-derivatized polyacrylamide supports, or conversely, bromoacetyl-oligonucleotides are immobilized on thiol-polyacrylamide supports.In a further aspect, this invention relates to bromoacetyl-oligonucleotides that may be immobilized on thiol-polyacrylamide solid supports, thiol-oligonucleotides immobilized on bromoacetyl-derivatized polyacrylamide supports as well as to methods for capture of nucleic acids by oligonucleotides attached to polyacrylamide solid supports, either by direct capture or in sandwich hybridization formats.

    Abstract translation: 本发明涉及将寡核苷酸共价连接到基本上在其5'端的固体支持物的方法和手段。 根据本发明，硫醇 - 寡核苷酸连接到溴乙酰基衍生的聚丙烯酰胺载体上，或者相反地，溴乙酰基寡核苷酸被固定在巯基 - 聚丙烯酰胺载体上。 在另一方面，本发明涉及可固定在硫醇 - 聚丙烯酰胺固体支持物上的溴乙酰基寡核苷酸，固定在溴乙酰基衍生的聚丙烯酰胺载体上的硫醇寡核苷酸以及通过与聚丙烯酰胺固体支持物连接的寡核苷酸捕获核酸的方法 ，通过直接捕获或夹心杂交形式。

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