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71. 发明公开

EP1674565A3 Method for enhancing enzyme activity at elevated temperature 失效
标题翻译：在升高的温度增加的酶活性的方法
公开(公告)号：EP1674565A3
公开(公告)日：2007-02-14
申请号：EP06007353.3
申请日：1997-07-24
申请人： THE INSTITUTE OF PHYSICAL & CHEMICAL RESEARCH , Hayashizaki, Yoshihide
发明人： Hayashizaki, Yoshihide
IPC分类号： C12N9/96 , C12N9/12 , C12N9/22 , C12Q1/68
CPC分类号： C12N9/1276 , C12N9/1252 , C12N9/22 , C12N9/96
摘要： A method for enhancing activity of enzyme at an elevated temperature which comprises adding a substance exhibiting chaperone function such as a saccharide to a reaction mixture containing the enzyme. The method can improve activity of enzymes more easily and more effectively and hence afford increased enzyme activity at an elevated temperature.

72. 发明公开

EP1018549A4 METHOD FOR ISOLATING DNA 有权
标题翻译：方法DNA分离。
公开(公告)号：EP1018549A4
公开(公告)日：2002-12-18
申请号：EP98943033
申请日：1998-09-18
申请人： RIKEN , HAYASHIZAKI YOSHIHIDE
发明人： HAYASHIZAKI YOSHIHIDE-INST PHY , CARNINCI PIERO-INST OF PHYS CH
IPC分类号： C12N15/09 , C07H21/04 , C12N15/10
CPC分类号： C12N15/1017 , C12N15/1006
摘要： A method for isolating DNA comprising: (A) feeding into a column provided with a DNA-binding carrier on a membrane filter a solution which contains a biological sample containing a salt, a cationic surfactant and a DNA and in which the concentration of the salt is not lower than the precipitation inhibition starting concentration and thus making the DNA contained in the biological sample to bind to the DNA-binding carrier; (B) sucking off the solution from the column and separating the carrier having the DNA bound thereto from other components; (C) feeding a DNA-dissociating solution into the column to thereby dissociate the DNA from the carrier; and (D) sucking off the dissociation solution from the column and taking up the solution containing the dissociated DNA. According to this method, a purified DNA can be taken up in a high yield without resort to any pretreatment of a biological sample.

73. 发明公开

EP1046648A4 3'-DEOXYRIBONUCLEOTIDE DERIVATIVES 失效
标题翻译： 3'-DESOXYRIBONUKLEOTID衍
公开(公告)号：EP1046648A4
公开(公告)日：2002-09-11
申请号：EP98929852
申请日：1998-07-06
申请人： RIKAGAKU KENKYUSHO , WAKO PURE CHEMICAL INDUSTIES L , HAYASHIZAKI YOSHIHIDE
发明人： HAYASHIZAKI YOSHIHIDE , OZAWA KAORI , FUJIO KAZUNARI , TANAKA TAKUMI
IPC分类号： C12N15/09 , C07H19/04 , C07H19/067 , C07H19/10 , C07H19/14 , C07H19/167 , C07H19/20 , C12Q1/68 , C07H21/00
CPC分类号： C07H19/04
摘要： 3'-Deoxyribonucleotide derivatives represented by the following general formula [I]: Q-V-(CH2)n-NR-R wherein Q represents a 3'-deoxyribonucleotide residue; n is an integer of 4 or more; V represents -CC- or -CH=CH-; and R represents a fluorescent group. Because of having been improved in the uptake efficiency by RNA polymerases, these derivatives are useful as terminators in DNA base sequencing methods with the use of RNA polymerases.

74. 发明公开

EP1046648A1 3'-DEOXYRIBONUCLEOTIDE DERIVATIVES 失效
标题翻译： 3'-DESOXYRIBONUKLEOTID衍
公开(公告)号：EP1046648A1
公开(公告)日：2000-10-25
申请号：EP98929852.6
申请日：1998-07-06
申请人： THE INSTITUTE OF PHYSICAL & CHEMICAL RESEARCH , Wako Pure Chemical Industies, Ltd. , Hayashizaki, Yoshihide
发明人： HAYASHIZAKI, Yoshihide , OZAWA, Kaori, Wako Pure Chemical Industries, Ltd. , FUJIO, Kazunari, Wako Pure Chemical Ind. Ltd. , TANAKA, Takumi, Wako Pure Chemical Industries, Ltd
IPC分类号： C07H19/10 , C07H19/14 , C07H19/20 , C12Q1/68
CPC分类号： C07H19/04
摘要： 3'-Deoxyribonucleotide derivatives represented by the following general formula [I]: Q-V-(CH 2 ) n -NR-R wherein Q represents a 3'-deoxyribonucleotide residue; n is an integer of 4 or more; V represents -C≡C- or -CH=CH-; and R represents a fluorescent group. Because of having been improved in the uptake efficiency by RNA polymerases, these derivatives are useful as terminators in DNA base sequencing methods with the use of RNA polymerases.
摘要翻译：式（I）的衍生物是新的：Q = 3'-脱氧核糖核苷酸残基; n => = 4; V = Ctriple boundC或CH = CH; R =荧光基团。

75. 发明公开

EP1006180A1 Method for producing double stranded DNA whose terminal homopolymer part ist eliminated and method for determining nucleotide sequence 有权
标题翻译：用于制备双链DNA，其终端部均聚物被消除，和方法，用于核苷酸序列的测定方法
公开(公告)号：EP1006180A1
公开(公告)日：2000-06-07
申请号：EP99122104.5
申请日：1999-11-05
申请人： Riken , Hayashizaki, Yoshihide
发明人： Hayashizaki, Yoshihide
IPC分类号： C12N15/10
CPC分类号： C12N15/1096 , Y10T436/143333
摘要： Disclosed is a method for producing double-stranded DNA comprising treating double-stranded DNA having a homopolymer part or parts at one or both ends with a restriction enzyme to partly or fully eliminate at least one of the homopolymer part or parts. The restriction enzyme is capable of cleaving double-stranded DNA at a cleavage site separate from a recognition site therefor. Disclosed is a method for determining a nucleotide sequence of double-stranded DNA utilizing one or both strands of the double-stranded DNA as a template, wherein the double-stranded DNA used as the template is double-stranded DNA prepared by the above method of the present invention. The present invention provides a method for producing double-stranded DNA a part or all of which homopolymer part, which may inhibit nucleotide sequence determination, is eliminated, and a method for determining a nucleotide sequence utilizing as a template the double-stranded DNA prepared by the above method of the present invention, whose homopolymer part is partially or fully eliminated.
摘要翻译：公开了一种用于产生双链DNA包括处理具有均聚物部分或多个部分的一个或用限制酶两端部分或完全消除均聚物部分或部分中的至少一个双链DNA的方法。限制性内切酶是能够在切割位点从一个识别位点分开为此切割双链DNA的。公开了用于确定性的采矿方法双链DNA利用一个或双链DNA作为模板的两条链，worin用作模板的双链DNA的核苷酸序列是由上述方法制备的双链DNA 了本发明。本发明提供一种制备双链DNA的一部分或全部均聚物部分，其可以抑制核苷酸序列测定的方法，被消除，以及用于确定性采矿方法的核苷酸序列利用作为模板通过制备的双链DNA 本发明中，其均聚物部分被部分或完全消除的上述方法。

76. 发明授权

EP0821059B1 Method for reverse transcription 失效
标题翻译：一种用于反转录方法
公开(公告)号：EP0821059B1
公开(公告)日：2005-12-21
申请号：EP97112672.7
申请日：1997-07-24
申请人： THE INSTITUTE OF PHYSICAL & CHEMICAL RESEARCH , Hayashizaki, Yoshihide
发明人： Hayashizaki, Yoshihide Tsukuba Life Science Center
IPC分类号： C12N9/12 , C12Q1/68 , C12N15/10 , C07H21/02
CPC分类号： C12N15/1096 , C12Q1/6844 , C12Q2527/101 , C12Q2521/107 , C12Q2527/125

77. 发明公开

EP1264899A3 Methods and kits for preparing and detecting RNA probes 审中-公开
标题翻译：方法和试剂盒用于制备和用于检测RNA探针的
公开(公告)号：EP1264899A3
公开(公告)日：2003-12-03
申请号：EP02012176.0
申请日：2002-06-03
申请人： Riken , Hayashizaki, Yoshihide
发明人： Hayashizaki, Yoshihide , Okazaki, Yasushi, c/o Riken Yokohama Institute
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6865 , C12Q2521/119 , C12Q2525/101 , C12Q2563/107
摘要： A method of preparing labeled RNA probe by reacting RNA polymerase in the presence of a DNA fragment comprising a promoter sequence for the RNA polymerase and substrates of the RNA polymerase. In the method, at least one of said substrates comprises said label, and said RNA polymerase is mutant RNA polymerase where at least one of the amino acids of wild type RNA polymerase has been modified to permit incorporation of the substrate having a label or to improve the incorporation of the substrate having a label. A method of detecting targeted nucleic acid in which targeted nucleic acid and labeled RNA probe prepared by the above method are mixed and RNA probe that has hybridized with the targeted nucleic acid is selectively detected. A kit for preparing labeled RNA probe comprising (1) RNA polymerase, (2) DNA comprising a promoter sequence for said RNA polymerase, (3) substrates of said RNA polymerase, and (4) optionally an instruction manual. In the kit, at least one of said substrates comprises a label and said RNA polymerase is mutant RNA polymerase where at least one of the amino acids of wild type RNA polymerase has been modified to permit incorporation of said substrate having a label or to improve the incorporation of said substrate having a label.

78. 发明公开

EP1197552A3 Method of preparing normalized and/or subtracted cDNA 有权
标题翻译：用于生产标准的和/或相减的cDNA的方法
公开(公告)号：EP1197552A3
公开(公告)日：2003-07-30
申请号：EP01120108.4
申请日：2001-08-22
申请人： Riken , Hayashizaki, Yoshihide
发明人： Hayashizaki, Joshihide
IPC分类号： C12N15/10 , C12Q1/68
CPC分类号： C12N15/1096
摘要： A method of preparing normalized and/or subtracted cDNA; a method in which the cDNA that is normalized and/or subtracted is in the form of uncloned cDNA (cDNA tester); a method of preparing normalized and/or subtracted cDNA comprising the steps of: (a) preparing cDNA tester; (b) preparing normalization and/or subtraction RNA driver; (c) conducting normalization and/or subtraction in two steps in any order, or conducting normalization/subtraction as a single step and mixing the normalization/subtraction RNA driver with said cDNA tester; (d) adding an enzyme capable of cleaving single strand sites on RNA drivers nonspecifically bound to cDNA tester; (e) removing said single strand RNA driver cleaved in step d) from the tester and removing tester/driver hybrids; and (f) recovering the normalized and/or subtracted cDNA; and a method of efficiently preparing normalized and/or subtracted long-chain, full-coding, and full-length cDNA libraries are provided.

79. 发明公开

EP0978569A4 METHOD OF DNA SEQUENCING 失效
标题翻译：法DNA测序
公开(公告)号：EP0978569A4
公开(公告)日：2002-09-04
申请号：EP98929853
申请日：1998-07-06
申请人： RIKAGAKU KENKYUSHO , HAYASHIZAKI YOSHIHIDE
发明人： HAYASHIZAKI YOSHIHIDE
IPC分类号： C12N15/09 , C12N1/21 , C12N15/54 , C12Q1/68 , C12N9/12 , C12P19/34
CPC分类号： C12Q1/6869 , C12Q2521/119
摘要： A method of DNA sequencing comprising reacting a ribonucleoside 5'-triphosphate with a 3'dNTP derivative in the presence of a mutated RNA polymerase modified so as to enhance the ability to take up the 3'dNTP derivative and a DNA fragment containing a promoter sequence for the RNA polymerase, separating the nucleic acid transcription product thus obtained, and reading the sequence of the nucleic acid from the fraction thus separated. By using this method, a long-chain transcription product can be formed and more accurate sequence data with little change in the signals from labeled deoxyribonucleotides can be obtained.

80. 发明公开

EP1197552A2 Method of preparing normalized and/or subtracted cDNA 有权
标题翻译： Methode zur Herstellung von genormten und / oder subtrahierten cDNA
公开(公告)号：EP1197552A2
公开(公告)日：2002-04-17
申请号：EP01120108.4
申请日：2001-08-22
申请人： Riken , Hayashizaki, Yoshihide
发明人： Hayashizaki, Joshihide
IPC分类号： C12N15/10 , C12Q1/68
CPC分类号： C12N15/1096
摘要： A method of preparing normalized and/or subtracted cDNA; a method in which the cDNA that is normalized and/or subtracted is in the form of uncloned cDNA (cDNA tester); a method of preparing normalized and/or subtracted cDNA comprising the steps of: (a)
preparing cDNA tester; (b) preparing normalization and/or subtraction RNA driver; (c) conducting normalization and/or subtraction in two steps in any order, or conducting normalization/subtraction as a single step and mixing the normalization/subtraction RNA driver with said cDNA tester; (d) adding an enzyme capable of cleaving single strand sites on RNA drivers nonspecifically bound to cDNA tester; (e) removing said single strand RNA driver cleaved in step d) from the tester and removing tester/driver hybrids; and (f) recovering the normalized and/or subtracted cDNA; and a method of efficiently preparing normalized and/or subtracted long-chain, full-coding, and full-length cDNA libraries are provided.
摘要翻译：制备标准化和/或减去的cDNA的方法; 标准化和/或减去的cDNA是未克隆的cDNA（cDNA测试仪）的形式的方法; 制备标准化和/或减去的cDNA的方法，包括以下步骤：（a>制备cDNA测试仪;（b）制备标准化和/或减法RNA驱动剂;（c）以任何顺序以两个步骤进行归一化和/或减法，或作为单步进行归一化/减法，并将归一化/减法RNA驱动剂与所述cDNA测试仪混合;（d）加入能够切割非特异性结合cDNA测试仪的RNA驱动单链位点的酶;（e）除去所述单链 RNA驱动器在步骤d）中从测试仪断开并去除测试仪/驱动器混合物; 和（f）回收标准化和/或减去的cDNA; 并提供了有效制备标准化和/或减去的长链，全编码和全长cDNA文库的方法。

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