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1. 发明专利

JP2014027955A Gene expression analysis method using two-dimensional cdna library 有权
标题翻译：使用二维CDNA库的基因表达分析方法
公开(公告)号：JP2014027955A
公开(公告)日：2014-02-13
申请号：JP2013234715
申请日：2013-11-13
申请人： Hitachi Ltd , 株式会社日立製作所
发明人： SHIRAI MASATAKA , KANBARA HIDEKI , TANIGUCHI KIYOMI
IPC分类号： C12M1/00 , C12Q1/68 , C40B40/08
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： PROBLEM TO BE SOLVED: To provide a method and/or means for picking out one cell and analyzing it and for quantitatively monitoring expression level of various genes with maintaining two-dimensional information of a tissue.SOLUTION: The invention provides a method for obtaining gene expression level in any loci or all of the target loci at one cell level positional resolution by producing cDNA library from mRNA with maintaining two-dimensional cell distribution information in a living tissue. More concretely, sheet shape cDNA library is produced from mRNA with maintaining two-dimensional cell distribution information in the living tissue and this is repeatedly used in gene expression detection steps, thereby realizing a high precision expression distribution measurement for a large number of genes.
摘要翻译：要解决的问题：提供一种选择一个细胞并进行分析的方法和/或方法，并且通过维持组织的二维信息来定量监测各种基因的表达水平。解决方案：本发明提供了获得基因的方法通过从维持生物体组织维持二维细胞分布信息的mRNA产生cDNA文库，在一个细胞水平位置分辨率下的任何位点或所有靶基因座中的表达水平。更具体地，由mRNA产生片状cDNA文库，维持活体组织中的二维细胞分布信息，并重复用于基因表达检测步骤，从而实现大量基因的高精度表达分布测定。

2. 发明专利

JP2009131176A Method for nucleic acid quantitation 有权
标题翻译：核酸定量方法
公开(公告)号：JP2009131176A
公开(公告)日：2009-06-18
申请号：JP2007309089
申请日：2007-11-29
申请人： Hitachi Ltd , 株式会社日立製作所
发明人： TANIGUCHI KIYOMI , KANBARA HIDEKI
IPC分类号： C12Q1/68 , C12N15/09
CPC分类号： C12Q1/6851 , C12Q2545/101 , C12Q2525/161
摘要： PROBLEM TO BE SOLVED: To provide a novel convenient approach for DNA quantitative analysis that overcomes the disadvantages of conventional formulations.
SOLUTION: A standard DNA sample is prepared by introducing a single-base substitution into target DNA, and a predetermined amount thereof is mixed with a target DNA sample. The target and standard DNAs are amplified using the same primers designed to amplify a region comprising the single-base substitution site. A probe capable of binding to a site immediately before the single-base substitution site, ddATP, ddGTP, ddCTP, and ddTTP are sequentially added one by one to perform a complementary strand synthesis reaction. Luciferase reaction-induced luminescence derived from the formed pyrophosphoric acid is detected. The target DNA is quantitated from the amount of the detected luminescence and the amount of the added standard DNA sample.
COPYRIGHT: (C)2009,JPO&INPIT
摘要翻译：要解决的问题：为了提供克服常规制剂的缺点的DNA定量分析的新颖方便的方法。解决方案：通过将靶向DNA中的单碱基取代引入，并将其预定量与目标DNA样品混合来制备标准DNA样品。使用设计用于扩增包含单碱基取代位点的区域的相同引物扩增靶标和标准DNA。依次向单碱基置换位点ddATP，ddGTP，ddCTP和ddTTP之前结合位点的探针依次加入进行互补链合成反应。检测到由形成的焦磷酸衍生的荧光素酶反应诱导的发光。从所检测的发光量和添加的标准DNA样品的量定量靶DNA。版权所有（C）2009，JPO＆INPIT

3. 发明专利

JP2006271311A Method for analyzing nucleic acid 审中-公开
标题翻译：分析核酸的方法
公开(公告)号：JP2006271311A
公开(公告)日：2006-10-12
申请号：JP2005098398
申请日：2005-03-30
申请人： Hitachi Ltd , 株式会社日立製作所
发明人： TANIGUCHI KIYOMI , NAGAI KEIICHI , MATSUNAGA HIROKO
IPC分类号： C12Q1/68 , C12N15/09 , C12Q1/66
CPC分类号： C12Q1/683 , C12Q2537/113 , C12Q2523/125 , C12Q2521/307
摘要： PROBLEM TO BE SOLVED: To provide a method for analyzing a nucleic acid, by which the presence or absence of a gene mutation and the methylated cytosine of a CpG dinucleotide contained in a target sequence originated from a test DNA can simply and highly accurately be detected.
SOLUTION: This method for analyzing the nucleic acid is characterized by cleaving the non-complemental strand sites of a double-stranded nucleic acid sample with a single-stranded specific endonuclease, hybridizing the obtained nucleic acid fragment with a probe containing a base sequence partially or wholly identical to either one strand of the double-stranded nucleic acid sample, subjecting the probe to an elongation reaction from the hybridized nucleic acid fragment, and then optically detecting pyrophosphoric acid produced by the elongation reaction to judge the presence or absence of the non-complemental strand sites in the double-stranded nucleic acid sample.
COPYRIGHT: (C)2007,JPO&INPIT
摘要翻译：待解决的问题：提供一种用于分析核酸的方法，通过该方法，源自测试DNA的靶序列中包含的CpG二核苷酸的基因突变和甲基化胞嘧啶的存在或不存在可以简单高度地准确检测。解决方案：用于分析核酸的方法的特征在于用单链特异性内切核酸酶切割双链核酸样品的非互补链位点，将获得的核酸片段与含有碱基的探针杂交序列部分或全部与双链核酸样品的一条链相同，对探针进行来自杂交核酸片段的延伸反应，然后光学检测由延伸反应产生的焦磷酸，以判断是否存在双链核酸样品中的非互补链位点。版权所有（C）2007，JPO＆INPIT

4. 发明专利

JP2006006274A Gene testing method 有权
标题翻译：基因测试方法
公开(公告)号：JP2006006274A
公开(公告)日：2006-01-12
申请号：JP2004191781
申请日：2004-06-29
申请人： Hitachi Ltd , 株式会社日立製作所
发明人： NAGAI KEIICHI , OKANO KAZUNOBU , NODA HIDEYUKI , MATSUNAGA HIROKO , TANIGUCHI KIYOMI , YAZAWA YOSHIAKI , KAJIYAMA TOMOHARU
IPC分类号： C12Q1/68 , C12N15/09 , G01N21/76 , G01N33/53 , G01N37/00
CPC分类号： C12Q1/6827 , C12Q1/6837 , C12Q2565/537 , C12Q2565/301 , C12Q2525/155
摘要： PROBLEM TO BE SOLVED: To provide a gene testing method capable of testing single nucleotide polymorphisms (SNPs) in a plurality of variation regions at a low cost by a simple operation and capable of realizing gene diagnosis on a clinical site. SOLUTION: This gene testing method comprises a process for hybridizing a sample nucleic acid having an anchor sequence at the 5' end with a support having a surface to which a probe containing a sequence complementary to a sequence (SNP region) to be detected is immobilized, a process for conducting elongation reaction of a complementary chain of the probe by using the sample nucleic acid as a template, a process for dissociating and removing the sample nucleic acid from the elongated chain of the probe synthesized in the elongation reaction, a process for conducting elongation reaction of a complementary chain with a primer having a sequence identical to the anchor sequence by using the elongated chain of the probe dissociated in the above as a template, and a process for detecting pyrophosphoric acid formed by the elongation reaction of the primer with biochemical luminescence, so that an SNP type of the sample nucleic acid is determined. COPYRIGHT: (C)2006,JPO&NCIPI
摘要翻译：要解决的问题：提供能够通过简单的操作以低成本测试多个变异区域中的单核苷酸多态性（SNP）的基因测试方法，并且能够在临床现场实现基因诊断。解决方案：该基因测试方法包括将具有5'端锚定序列的样品核酸与具有表面的载体杂交的方法，所述载体具有包含与序列互补序列（SNP区）的探针的表面检测到的固定化物，通过使用样品核酸作为模板进行探针的互补链的伸长反应的方法，从延伸反应合成的探针的细长链中解离和除去样品核酸的方法，通过使用在上述中解离的探针的细长链作为模板，使互补链与具有与锚定序列相同的序列的引物进行延伸反应的方法，以及通过延伸反应形成的焦磷酸的检测方法该引物具有生化发光，从而确定样品核酸的SNP型。版权所有（C）2006，JPO＆NCIPI

5. 发明专利

JP2011147415A Method for uniform amplification of nucleic acid and highly sensitive detection method 有权
标题翻译：用于均匀放大核酸和高敏感性检测方法的方法
公开(公告)号：JP2011147415A
公开(公告)日：2011-08-04
申请号：JP2010013159
申请日：2010-01-25
申请人： Hitachi Ltd , 株式会社日立製作所
发明人： KANBARA HIDEKI , MATSUNAGA HIROKO , TANIGUCHI KIYOMI
IPC分类号： C12N15/09 , C12Q1/68
CPC分类号： C12Q1/686 , C12Q2525/151 , C12Q2531/125 , C12Q2537/143
摘要： PROBLEM TO BE SOLVED: To provide a method for rapidly and readily amplifying a small amount of nucleic acids in high amplification factor. SOLUTION: The method for amplification of the nucleic acids includes a step for introducing a repeated sequence containing an already-known sequence to the 3' terminus of a target nucleic acid chain containing a target sequence or a complementary sequence thereof, a step for hybridizing a plurality of oligonucleotides hybridizable with the repeated sequence, with the repeated sequence introduced into the target nucleic acid chain, and a step for carrying out the synthesis of a complementary chain using the target nucleic acid chain as a template by using the oligonucleotide as a primer to form a plurality of the nucleic acid-complementary chains from a single target nucleic acid chain. COPYRIGHT: (C)2011,JPO&INPIT
摘要翻译：待解决的问题：提供以高放大因子快速且容易地扩增少量核酸的方法。解决方案：扩增核酸的方法包括将包含已知序列的重复序列引入含有靶序列或其互补序列的靶核酸链的3'末端的步骤，步骤用于将可重复序列杂交的多个寡核苷酸与引入靶核酸链的重复序列杂交，以及使用寡核苷酸作为模板，使用靶核酸链作为模板进行互补链的合成的步骤从单个靶核酸链形成多个核酸互补链的引物。版权所有（C）2011，JPO＆INPIT

6. 发明专利

JP2007319028A Method for determining gene expression of single cell 有权
标题翻译：用于确定单细胞基因表达的方法
公开(公告)号：JP2007319028A
公开(公告)日：2007-12-13
申请号：JP2006150189
申请日：2006-05-30
申请人： Hitachi Ltd , 株式会社日立製作所
发明人： TANIGUCHI KIYOMI , KANBARA HIDEKI , KAJIYAMA TOMOHARU
IPC分类号： C12Q1/68 , C12N15/09
CPC分类号： C12Q1/6811 , C12N15/1096 , C12Q1/6809 , C12Q1/6834 , C12Q2565/537 , C12Q2525/173 , C12Q2521/301 , C12Q2561/113
摘要： PROBLEM TO BE SOLVED: To provide a method for suitably and quantitatively analyzing the gene expression of a single cell. SOLUTION: This method for detecting nucleic acid is provided by comprising a process of collecting the single cell from a specimen containing at least the single cell, a process of lysing the cell for dissolving the cell membrane of the collected single cell to elute the nucleic acids of the cell, a DNA-decomposing process of decomposing the DNA from the eluted nucleic acids with a DNA-decomposing enzyme, a process of hybridizing mRNA among RNAs contained in the single cell with an oligo(dT) immobilized on a carrier, a process of performing a reverse transcription reaction on the mRNA hybridized with the oligo(dT) to prepare a single cell-derived cDNA-immobilized carrier library consisting of the single cell-derived cDNA immobilized on the carrier and a process of detecting the amplified amount while performing an amplification reaction of the cDNA immobilized on the carrier. COPYRIGHT: (C)2008,JPO&INPIT
摘要翻译：待解决的问题：提供适当和定量分析单细胞的基因表达的方法。解决方案：这种检测核酸的方法是通过包括从至少含有单个细胞的样品中收集单个细胞的方法来提供的，该方法是将收集的单个细胞的细胞膜溶解的细胞裂解以洗脱细胞的核酸，用DNA分解酶分解来自洗脱的核酸的DNA的DNA分解过程，将单细胞中包含的RNA中的mRNA与固定在载体上的寡核苷酸（dT）杂交的过程，对与寡聚（dT）杂交的mRNA进行逆转录反应的方法，以制备由固定在载体上的单细胞衍生的cDNA组成的单细胞衍生的cDNA固定的载体文库，以及检测扩增的同时进行固定在载体上的cDNA的扩增反应。版权所有（C）2008，JPO＆INPIT

7. 发明专利

DE602007004771D1 未知
公开(公告)号：DE602007004771D1
公开(公告)日：2010-04-01
申请号：DE602007004771
申请日：2007-05-03
申请人： HITACHI LTD
发明人： TANIGUCHI KIYOMI , KAMBARA HIDEKI , KAJIYAMA TOMOHARU
IPC分类号： C12N15/10 , C12Q1/68

8. 发明专利

DE602007012766D1 Verfahren zur quantitativen cDNA-Analyse in Einzelzellen 未知
公开(公告)号：DE602007012766D1
公开(公告)日：2011-04-07
申请号：DE602007012766
申请日：2007-05-03
申请人： HITACHI LTD
发明人： TANIGUCHI KIYOMI , KAMBARA HIDEKI , KAJIYAMA TOMOHARU
IPC分类号： C12N15/10 , C12Q1/68

9. 发明公开

EP2881465A4 TAG-SEQUENCE-ATTACHED TWO-DIMENSIONAL CDNA LIBRARY DEVICE, AND GENE EXPRESSION ANALYSIS METHOD AND GENE EXPRESSION ANALYSIS APPARATUS EACH UTILIZING SAME 有权
标题翻译：一个标签序列连接有两个维CDNS库和基因表达分析方法和基因表达分析设备
公开(公告)号：EP2881465A4
公开(公告)日：2016-04-20
申请号：EP12882489
申请日：2012-07-30
申请人： HITACHI LTD
发明人： SHIRAI MASATAKA , KAMBARA HIDEKI , TANIGUCHI KIYOMI , TANABE MAIKO
IPC分类号： C12N15/09 , C12M1/00 , C12N15/10 , C12Q1/68
CPC分类号： C12Q1/6837 , C12N15/1096 , C12Q2525/155 , C12Q2565/537 , C12Q2565/601
摘要： The present invention relates to a method, a device, and an apparatus for analyzing the expression of a gene in single cells. Specifically, the present invention relates to: a device for gene expression analysis, characterized by including a support, in which a nucleic acid probe having a test nucleic acid capture sequence and a known sequence, and further containing a cell recognition tag sequence which differs depending on the difference in position on the surface of the support or in the vicinity of the surface thereof, and a common primer sequence having a known sequence is two-dimensionally distributed and immobilized on the surface of the support or in the vicinity of the surface thereof; and a method and an apparatus using the device for gene expression analysis.
摘要翻译：本发明涉及一种方法，设备，以及装置，用于分析单个细胞中的基因的表达。具体而言，本发明涉及：用于基因表达分析的装置，通过包括支持体，其中具有测试核酸捕获序列和已知的序列的核酸探针，和进一步含有不同，取决于小区识别标签序列为特征的在该支持体或在其表面的附近的表面，并具有已知序列的一个共同的引物序列上的位置差是二维分散和固定在载体上的表面上或在表面附近或其 ; 和的方法和使用该装置用于基因表达分析的装置。

10. 发明公开

EP2975123A4 TWO-DIMENSIONAL CELL ARRAY DEVICE AND APPARATUS FOR GENE QUANTIFICATION AND SEQUENCE ANALYSIS 有权
标题翻译：二维细胞基质设备和设备进行基因定量和序列分析
公开(公告)号：EP2975123A4
公开(公告)日：2016-11-30
申请号：EP13878277
申请日：2013-03-12
申请人： HITACHI LTD
发明人： SHIRAI MASATAKA , KAMBARA HIDEKI , TANIGUCHI KIYOMI , TANABE MAIKO
IPC分类号： C12N15/10 , C12M1/00 , C12Q1/68
CPC分类号： C12N15/1003 , B01L3/502761 , B01L7/52 , B01L2200/0631 , B01L2200/0668 , B01L2200/10 , B01L2300/0636 , B01L2300/0819 , B01L2300/0851 , B01L2300/0864 , B01L2300/1805 , B01L2400/0421 , C12N15/1096 , C12Q1/6874 , C12Q2539/10
摘要： In order to conduct gene expression analysis of a number of genes in a number of cells, it has been necessary to separate cells, extract genes therefrom, amplify nucleic acids, and perform sequence analysis. However, separation of cells imposes damages on the cells, and it requires the use of an expensive system. Gene expression analysis in each cell can be carried out with high accuracy by arranging a pair of structures comprising a cell trapping section and a nucleic acid trapping section in a vertical direction to extract individual genes in relevant cells, synthesizing cDNA in the nucleic acid trapping section, amplifying nucleic acids, and analyzing the sequences using a next-generation sequencer.

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