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    METHODS FOR NUCLEIC ACID ASSEMBLY AND HIGH THROUGHPUT SEQUENCING
    4.
    发明公开
    METHODS FOR NUCLEIC ACID ASSEMBLY AND HIGH THROUGHPUT SEQUENCING 审中-公开

    公开(公告)号：EP3483311A1

    公开(公告)日：2019-05-15

    申请号：EP18194361.4

    申请日：2013-06-24

    申请人： Gen9, Inc.

    发明人： HUDSON, Michael, E. ,  KUNG, Li-yun A. ,  SCHINDLER, Daniel ,  ARCHER, Stephen ,  SAAEM, Ishtiaq

    IPC分类号： C40B50/06 ,  C12N15/10 ,  C12N15/66

    摘要： Methods and apparatus of some aspects of the invention relate to the synthesis of high fidelity polynucleotides. In particular, aspects of the invention relate to concurrent enzymatic removal of amplification sequences and ligation of processed oligonucleotides into nucleic acid assemblies. According to some embodiments, the invention provides a method for producing a target nucleic acid having a predefined sequence. In some embodiments, the method comprises the step of providing a plurality of oligonucleotides, wherein each oligonucleotides comprises (i) an internal sequence identical to a different portion of a sequence of a target nucleic acid, (ii) a 5' sequence flanking the 5' end of the internal sequence and a 3' flanking sequence flanking the 3' end of the internal sequence, each of the flanking sequence comprising a primer recognition site for a primer pair and a restriction enzyme recognition site.

    COMPOSITIONS AND METHODS FOR HIGH FIDELITY ASSEMBLY OF NUCLEIC ACIDS
    8.
    发明公开
    COMPOSITIONS AND METHODS FOR HIGH FIDELITY ASSEMBLY OF NUCLEIC ACIDS 审中-公开
    ZUSAMMENSETZUNGEN UND VERFAHREN ZUR HOCHQUALITATIVEN ANORDNUNG VONNUKLEINSÄUREN

    公开(公告)号：EP2944693A1

    公开(公告)日：2015-11-18

    申请号：EP15159715.0

    申请日：2012-08-23

    申请人： Gen9, Inc.

    发明人： Jacobson, Joseph ,  Schindler, Daniel ,  Lawton, Scott S.

    IPC分类号： C12N15/10 ,  C12N15/66 ,  C12P19/34 ,  C40B50/06 ,  G06F19/10

    CPC分类号： C12N15/66 ,  C12N15/10 ,  C12N15/1027 ,  C12N15/1031 ,  C12N15/1089 ,  C12P19/34 ,  C12Q2521/301 ,  C12Q2521/501 ,  G06F19/22

    摘要： Aspects of the invention relate to methods, compositions and algorithms for designing and producing a target nucleic acid. The method can include: (1) providing a plurality of blunt-end double-stranded nucleic acid fragments having a restriction enzyme recognition sequence at both ends thereof; (2) producing via enzymatic digestion a plurality of cohesive-end double-stranded nucleic acid fragments each having two different and non-complementary overhangs; (3) ligating the plurality of cohesive-end double-stranded nucleic acid fragments with a ligase; and (4) forming a linear arrangement of the plurality of cohesive-end double-stranded nucleic acid fragments, wherein the unique arrangement comprises the target nucleic acid. In certain embodiments, the plurality of blunt-end double-stranded nucleic acid fragments can be provided by: releasing a plurality of oligonucleotides synthesized on a solid support; and synthesizing complementary strands of the plurality of oligonucleotides using a polymerase based reaction.

    摘要翻译： 本发明的方面涉及用于设计和产生靶核酸的方法，组合物和算法。 该方法可以包括：（1）提供多个在其两端具有限制酶识别序列的平末端双链核酸片段; （2）通过酶消化产生多个具有两个不同和非互补突出端的内聚端双链核酸片段; （3）用连接酶连接多个内聚端双链核酸片段; 和（4）形成多个内聚端双链核酸片段的线性排列，其中独特的排列包含靶核酸。 在某些实施方案中，多个平末端双链核酸片段可以通过以下方式提供：释放在固体支持物上合成的多种寡核苷酸; 并使用基于聚合酶的反应合成多个寡核苷酸的互补链。