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    Apparatus for the separation and fractionation of differentially expressed gene fragments
    1.
    发明授权
    Apparatus for the separation and fractionation of differentially expressed gene fragments 失效
    用于分离和分离差异表达基因片段的装置

    公开(公告)号：US06387235B1

    公开(公告)日：2002-05-14

    申请号：US09385420

    申请日：1999-08-30

    申请人： Takashi Irie ,  Hideki Hasegawa ,  Hideki Kambara ,  Ken Ninomiya

    发明人： Takashi Irie ,  Hideki Hasegawa ,  Hideki Kambara ,  Ken Ninomiya

    IPC分类号： G01N2726

    CPC分类号： G01N27/44717

    摘要： An apparatus for the separation and fractionation of differentially expressed gene fragments includes a separating means including a capillary filled with a separation medium to separate DNA fragments by electrophoresis, a sampling means including sampling vessels to fractionate and sample the separated DNA fragments, according to their size, a transferring means to transfer buffer solution containing the separated DNA fragments to the sampling means, and a control means to control the sampling means based on a signal gained by detecting means, wherein a voltage for the electrophoresis and a length of the capillary are adjusted such that a spread in time of the separated DNA fragments caused by the transferring means during the transfer of the separated DNA fragments to the sampling vessels is smaller than a difference in separation time of the DNA fragments in the separating means.

    摘要翻译： 用于分离和分离差异表达的基因片段的装置包括分离装置，其包括填充有分离培养基的毛细管，以通过电泳分离DNA片段;采样装置，包括取样容器，以分离和样品分离的DNA片段，根据其大小 将含有分离的DNA片段的缓冲溶液转移到采样装置的转移装置，以及控制装置，用于根据由检测装置获得的信号控制采样装置，其中调整电泳电压和毛细管长度 使得在分离的DNA片段转移到采样容器期间由转移装置引起的分离的DNA片段的时间扩散小于分离装置中的DNA片段的分离时间的差异。

    DNA analysis apparatus
    4.
    发明授权
    DNA analysis apparatus 有权
    DNA分析仪器

    公开(公告)号：US08975066B2

    公开(公告)日：2015-03-10

    申请号：US12419399

    申请日：2009-04-07

    申请人： Hideki Kambara ,  Akihiko Kishimoto

    发明人： Hideki Kambara ,  Akihiko Kishimoto

    IPC分类号： C12M1/34 ,  C12M3/00 ,  C12Q1/68 ,  B01L7/00

    CPC分类号： C12Q1/6869 ,  B01J2219/00659 ,  B01J2219/00722 ,  B01L7/52 ,  C12Q2565/301 ,  C12Q2563/103 ,  C12Q2535/101

    摘要： Accurate and sensitive sequencing in pyrosequencing is achieved by allowing complementary strand synthesis reaction to proceed homogeneously and completely in a short time while performing luminescence reaction for a sufficiently long time. DNA as a sequencing target is immobilized on the surface of a solid support. Nucleic acid substrates are injected from a dispenser to the support site where complementary strand synthesis is in turn performed rapidly and completely in a short time under a small reaction volume. Next, the support together with the product thereon is moved into a luminescence reaction solution where luminescence reaction is in turn performed. Thus, a DNA complementary strand synthesis reaction site and a luminescence reaction site are completely separated. The support surface is also washed by dipping the support in the luminescence reaction solution that contains a luminescence reagent and an enzyme that degrades redundant nucleic acid substrates.

    摘要翻译： 焦磷酸测序中的精确和灵敏的测序通过使互补链合成反应在短时间内均匀且完全地进行，同时进行发光反应足够长的时间来实现。 作为测序靶的DNA被固定在固体支持物的表面上。 将核酸底物从分配器注射到支持部位，其中互补链合成在短时间内在小反应体积下快速且完全地进行。 接下来，将载体与其上的产物一起移动到进行发光反应的发光反应溶液中。 因此，DNA互补链合成反应位点和发光反应位点完全分离。 通过将载体浸渍在含有发光试剂的发光反应溶液和降解冗余核酸底物的酶中也可以洗涤载体表面。

    GENE EXPRESSION ANALYSIS METHOD USING TWO DIMENSIONAL cDNA LIBRARY
    5.
    发明申请
    GENE EXPRESSION ANALYSIS METHOD USING TWO DIMENSIONAL cDNA LIBRARY 审中-公开
    使用两维cDNA文库的基因表达分析方法

    公开(公告)号：US20120245053A1

    公开(公告)日：2012-09-27

    申请号：US13513605

    申请日：2010-11-29

    申请人： Masataka Shirai ,  Hideki Kambara ,  Kiyomi Taniguchi

    发明人： Masataka Shirai ,  Hideki Kambara ,  Kiyomi Taniguchi

    IPC分类号： C40B30/04

    CPC分类号： C12N15/1093 ,  C12Q1/6809 ,  C12Q1/6837 ,  C40B40/08 ,  C40B50/06 ,  G01N33/54306 ,  G01N2458/10 ,  C12Q2531/125 ,  C12Q2533/107 ,  C12Q2565/537

    摘要： The present invention provides a method and/or means for collecting and analyzing an individual cell in a tissue, and at the same time, quantitatively monitoring the expression levels of various genes while keeping two-dimensional information in the tissue. Specifically, the present invention provides a method comprising preparing a cDNA library from mRNA while keeping two-dimensional cellular distribution information and obtaining the gene expression levels at any site or all sites at a level of single cell. More specifically, the present invention provides a method comprising preparing a cDNA library in a sheet-form from mRNA while keeping two-dimensional cellular distribution information and repeatedly using the cDNA library in the detection of the gene expression, thereby allowing measurement of the expression distribution for a number of genes at a high accuracy.

    摘要翻译： 本发明提供了一种用于收集和分析组织中的单个细胞的方法和/或方法，并且同时定量监测各种基因的表达水平，同时保持组织中的二维信息。 具体地说，本发明提供一种方法，其包括从mRNA制备cDNA文库，同时保持二维细胞分布信息，并在单细胞水平的任何位点或所有位点获得基因表达水平。 更具体地，本发明提供了一种方法，其包括从mRNA制备片段的cDNA文库，同时保持二维细胞分布信息，并在检测基因表达中重复使用cDNA文库，从而允许测量表达分布 对于许多基因在高精度。

    Nucleic acid amplification device
    6.
    发明申请
    Nucleic acid amplification device 有权
    核酸扩增装置

    公开(公告)号：US20090215162A1

    公开(公告)日：2009-08-27

    申请号：US12292167

    申请日：2008-11-13

    申请人： Tomoharu Kajiyama ,  Masataka Shirai ,  Hideki Kambara

    发明人： Tomoharu Kajiyama ,  Masataka Shirai ,  Hideki Kambara

    IPC分类号： C12M1/40

    CPC分类号： B01L3/50851 ,  B01L3/5025 ,  B01L3/50857 ,  B01L2200/0668 ,  B01L2300/0819

    摘要： This invention provides a nucleic acid amplification device whereby the abundance of a target molecule can be maintained before and after a step of separately amplifying a sample such that highly accurate analysis results that can be applied to gene expression analysis can be obtained. Also, a nucleic acid amplification device having a structure in which a plurality of minute reaction cells each comprising a set of a bead-retaining space capable of retaining a single analysis bead and a reagent reaction space retaining no bead but having a volume that is large enough to cause a reagent reaction therein are positioned so as to form a planar face is provided.

    摘要翻译： 本发明提供了一种核酸扩增装置，其可以在分别扩增样品的步骤之前和之后维持目标分子的丰度，使得可以获得可应用于基因表达分析的高精度分析结果。 另外，具有这样的结构的核酸扩增装置，其中多个微小反应单元各自包含一组能够保留单个分析珠的珠保留空间和不保留珠而具有大体积的试剂反应空间 足以使其中的试剂反应定位成形成平面。

    Large-scale parallel nucleic acid analysis method
    7.
    发明申请
    Large-scale parallel nucleic acid analysis method 审中-公开
    大规模并行核酸分析方法

    公开(公告)号：US20080318244A1

    公开(公告)日：2008-12-25

    申请号：US12213448

    申请日：2008-06-19

    申请人： Hiroko Matsunaga ,  Hideki Kambara ,  Tomoharu Kajiyama

    发明人： Hiroko Matsunaga ,  Hideki Kambara ,  Tomoharu Kajiyama

    IPC分类号： C12Q1/68

    CPC分类号： C12Q1/6834 ,  C12Q1/6844 ,  C12Q2537/143 ,  C12Q2525/191 ,  C12Q2533/101 ,  C12Q2565/537

    摘要： It is intended to provide a technique for amplifying, individually and in parallel, nucleic acids contained in a mixture of plural kinds of nucleic acid samples. The present invention provides a nucleic acid analysis method comprising amplification means, whereby amplification reaction is performed in a reaction solution comprising a homogeneous solvent and comprising at least plural template nucleic acids and solid phase carriers comprising one or more kinds of amplification probes immobilized on the surface, to prevent amplified products attributed to two or more template nucleic acids from being replicated in one solid phase carrier. According to the present invention, plural kinds of analyte nucleic acid samples in a mixed state can be amplified individually and in parallel. This method achieves one solid phase carrier-one nucleic acid. Therefore, a higher density of solid phase carriers with obtained amplified products is easily achieved, leading to improved throughput of amplified product analysis. Reactions in all the amplification reaction steps are performed under homogeneous solvent conditions. Therefore, the method of the present invention is performed by convenient procedures and as such, is suitable to automation.

    摘要翻译： 旨在提供用于扩增多种核酸样品的混合物中包含的核酸的单独和并行扩增的技术。 本发明提供了包含扩增方法的核酸分析方法，其中扩增反应在包含均相溶剂并包含至少多个模板核酸的反应溶液中进行，并且包含固定在表面上的一种或多种扩增探针的固相载体 ，以防止归因于两种或更多种模板核酸的扩增产物被复制在一种固相载体中。 根据本发明，可以单独并行地扩增处于混合状态的多种分析物核酸样品。 该方法实现了一种固相载体一核酸。 因此，容易获得具有获得的扩增产物的较高密度的固相载体，从而提高扩增产物分析的产量。 所有扩增反应步骤中的反应在均相溶剂条件下进行。 因此，本发明的方法通过方便的程序进行，因此适用于自动化。

    Method and apparatus for sample preparation
    8.
    发明申请
    Method and apparatus for sample preparation 审中-公开
    样品制备方法和装置

    公开(公告)号：US20080241841A1

    公开(公告)日：2008-10-02

    申请号：US11984065

    申请日：2007-11-13

    申请人： Katsuji Murakawa ,  Sumiyo Takiguchi ,  Hideki Kambara

    发明人： Katsuji Murakawa ,  Sumiyo Takiguchi ,  Hideki Kambara

    IPC分类号： C12Q1/68 ,  C12P21/04 ,  C12M1/34

    CPC分类号： C12Q1/6806 ,  C12Q1/6851 ,  C12Q2527/143 ,  C12Q2563/173

    摘要： A method of the present invention comprises fractionating a sample solution containing analyte DNA molecules into small droplets, wherein the number M of the droplets is greater than the total number N of the DNA molecules, subjecting an emulsion containing the droplets to, for example, PCR amplification, and detecting the presence or absence (amount) of an amplicon obtained in each droplet by fluorescent detection using an intercalator or the like.

    摘要翻译： 本发明的方法包括将含有分析物DNA分子的样品溶液分级成小液滴，其中液滴的数量M大于DNA分子的总数N，使含有液滴的乳液经受例如PCR 通过使用嵌入剂等进行荧光检测，检测各液滴中获得的扩增子的存在或不存在（量）。