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    Addition of an adaptor by invasive cleavage
    1.
    发明授权
    Addition of an adaptor by invasive cleavage 有权
    通过侵入性切割添加适配器

    公开(公告)号：US09365896B2

    公开(公告)日：2016-06-14

    申请号：US13963723

    申请日：2013-08-09

    Applicant: Agilent Technologies, Inc.

    Inventor： Fredrik Dahl ,  Olof John Ericsson ,  Henrik Johansson ,  Maithreyan Srinivasan

    IPC: C12Q1/68

    CPC classification number: C12Q1/6855 ,  C12Q1/6806 ,  C12Q2525/101 ,  C12Q2525/179 ,  C12Q2521/307 ,  C12Q2525/191 ,  C12Q2525/307 ,  C12Q2531/131

    Abstract: This disclosure provides a method for adding an adaptor to a genomic sequence by invasive cleavage, as well as a kit for performing the method. In some embodiments, the method comprises: a) hybridizing genomic DNA to an adaptor comprising a double stranded region and a single stranded region comprising a 5′ overhang to produce a substrate for a flap endonuclease; b) cleaving the substrate using the flap endonuclease; c) ligating the recessed end of the double stranded region to the fragment to produce an adaptor-ligated DNA; d) intramolecularly ligating the ends of the adaptor-ligated DNA to produce a circular DNA molecule; and e) enzymatically processing the circular DNA molecule using an oligonucleotide that hybridizes to the adaptor and an enzyme. A kit for performing the method is also provided.

    Abstract translation: 本公开提供了通过侵入性切割将衔接子加入基因组序列的方法，以及用于进行该方法的试剂盒。 在一些实施方案中，所述方法包括：a）将基因组DNA与包含双链区域和包含5'突出端的单链区域的衔接子杂交以产生用于片段内切核酸酶的底物; b）使用翼片内切酶切割底物; c）将双链区域的凹入端连接到片段以产生衔接子连接的DNA; d）分子内连接衔接物连接的DNA的末端以产生环状DNA分子; 和e）使用与衔接子和酶杂交的寡核苷酸对环状DNA分子进行酶处理。 还提供了用于执行该方法的套件。

    ADDITION OF AN ADAPTOR BY INVASIVE CLEAVAGE
    5.
    发明申请
    ADDITION OF AN ADAPTOR BY INVASIVE CLEAVAGE 有权
    通过入侵清除添加适配器

    公开(公告)号：US20140113296A1

    公开(公告)日：2014-04-24

    申请号：US13963723

    申请日：2013-08-09

    Applicant: Agilent Technologies, Inc.

    Inventor： Fredrik Dahl ,  Olof John Ericsson ,  Henrik Johansson ,  Maithreyan Srinivasan

    IPC: C12Q1/68

    CPC classification number: C12Q1/6855 ,  C12Q1/6806 ,  C12Q2525/101 ,  C12Q2525/179 ,  C12Q2521/307 ,  C12Q2525/191 ,  C12Q2525/307 ,  C12Q2531/131

    Abstract: This disclosure provides method for adding an adaptor to a genomic sequence by invasive cleavage, as well as a kit for performing the method. In some embodiments, the method comprises: a) hybridizing genomic DNA to an adaptor comprising a double stranded region and a single stranded region comprising a 5′ overhang to produce a substrate for a flap endonuclease; b) cleaving the substrate using the flap endonuclease; c) ligating the recessed end of the double stranded region to the fragment to produce an adaptor-ligated DNA; d) intramolecularly ligating the ends of the adaptor-ligated DNA to produce a circular DNA molecule; and e) enzymatically processing the circular DNA molecule using an oligonucleotide that hybridizes to the adaptor and an enzyme. A kit for performing the method is also provided.

    Abstract translation: 本公开提供了通过侵入性切割将衔接子加入基因组序列的方法，以及用于进行该方法的试剂盒。 在一些实施方案中，所述方法包括：a）将基因组DNA与包含双链区域和包含5'突出端的单链区域的衔接子杂交以产生用于片段内切核酸酶的底物; b）使用翼片内切酶切割底物; c）将双链区域的凹入端连接到片段以产生衔接子连接的DNA; d）分子内连接衔接物连接的DNA的末端以产生环状DNA分子; 和e）使用与衔接子和酶杂交的寡核苷酸对环状DNA分子进行酶处理。 还提供了用于执行该方法的套件。

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