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公开(公告)号:US20130065281A1
公开(公告)日:2013-03-14
申请号:US13697665
申请日:2011-05-09
申请人: Yuko Nakabayashi , Takashi Uemori , Hiroyuki Mukai , Kiyozo Asada
发明人: Yuko Nakabayashi , Takashi Uemori , Hiroyuki Mukai , Kiyozo Asada
CPC分类号: C12N15/1096 , C12P19/34
摘要: A method for synthesizing cDNA characterized by preparing a reaction solution that does not allow an endodeoxyribonuclease to show its activity, without thermal deactivation of the endodeoxyribonuclease or removal of the endodeoxyribonuclease, and carrying out a reverse transcription reaction, wherein the reaction solution contains a treated sample and a reverse transcriptase, the treated sample being formed by treating a sample comprising RNA and DNA with the endodeoxyribonuclease to degrade DNA in the sample. The method and the kit for synthesizing cDNA of the present invention are widely useful in genetic engineering fields.
摘要翻译: 一种合成cDNA的方法,其特征在于制备不允许内脱氧核糖核酸酶显示其活性的反应溶液,不脱氧内脱氧核糖核酸酶或去除内切脱氧核糖核酸酶,并进行逆转录反应,其中反应溶液含有处理样品 和逆转录酶,通过用内切核糖核酸酶处理包含RNA和DNA的样品来降解样品中的DNA来形成经处理的样品。 用于合成本发明的cDNA的方法和试剂盒可广泛用于遗传工程领域。
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公开(公告)号:US09169482B2
公开(公告)日:2015-10-27
申请号:US13697665
申请日:2011-05-09
申请人: Yuko Nakabayashi , Takashi Uemori , Hiroyuki Mukai , Kiyozo Asada
发明人: Yuko Nakabayashi , Takashi Uemori , Hiroyuki Mukai , Kiyozo Asada
CPC分类号: C12N15/1096 , C12P19/34
摘要: A method for synthesizing cDNA characterized by preparing a reaction solution that does not allow an endodeoxyribonuclease to show its activity, without thermal deactivation of the endodeoxyribonuclease or removal of the endodeoxyribonuclease, and carrying out a reverse transcription reaction, wherein the reaction solution contains a treated sample and a reverse transcriptase, the treated sample being formed by treating a sample comprising RNA and DNA with the endodeoxyribonuclease to degrade DNA in the sample. The method and the kit for synthesizing cDNA of the present invention are widely useful in genetic engineering fields.
摘要翻译: 一种合成cDNA的方法,其特征在于制备不允许内脱氧核糖核酸酶显示其活性的反应溶液,不脱氧内脱氧核糖核酸酶或去除内切脱氧核糖核酸酶,并进行逆转录反应,其中反应溶液含有处理样品 和逆转录酶,通过用内切核糖核酸酶处理包含RNA和DNA的样品来降解样品中的DNA来形成经处理的样品。 用于合成本发明的cDNA的方法和试剂盒可广泛用于遗传工程领域。
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公开(公告)号:US20130295576A1
公开(公告)日:2013-11-07
申请号:US13980970
申请日:2012-01-20
申请人: Junko Yamamoto , Keiko Kubo , Takashi Uemori , Hiroyuki Mukai , Kiyozo Asada
发明人: Junko Yamamoto , Keiko Kubo , Takashi Uemori , Hiroyuki Mukai , Kiyozo Asada
IPC分类号: C12Q1/68
CPC分类号: C12N15/01 , C12Q1/6848 , C12Q1/686 , C12Q1/689 , C12Q2523/101 , C12Q2525/117
摘要: The present invention pertains to: a method for modifying a nucleic acid contained in a sample, said method including a step for bringing the sample into contact with a nucleic acid-modifying agent in the presence of an acidic polysaccharide and/or a nucleotide; and a method for selectively detecting a nucleic acid derived from living cells contained in the sample, said method including the following steps: (a) a step for modifying a nucleic acid contained in a sample according to the method for modifying a nucleic acid contained in a sample, which includes a step for bringing the sample into contact with a nucleic acid-modifying agent in the presence of an acidic polysaccharide and/or a nucleotide; and (b) a step for selectively detecting an unmodified nucleic acid from the sample after step (a). The present invention further pertains to a kit and composition for use in these methods.
摘要翻译: 本发明涉及:用于修饰样品中所含核酸的方法,所述方法包括在酸性多糖和/或核苷酸存在下使样品与核酸改性剂接触的步骤; 以及选择性地检测来自样品中含有的活细胞的核酸的方法,所述方法包括以下步骤:(a)根据包含在所述样品中的核酸的方法修饰样品中的核酸的步骤 样品,其包括在酸性多糖和/或核苷酸的存在下使样品与核酸改性剂接触的步骤; 和(b)在步骤(a)之后从样品中选择性地检测未修饰的核酸的步骤。 本发明还涉及用于这些方法的试剂盒和组合物。
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公开(公告)号:US08367328B2
公开(公告)日:2013-02-05
申请号:US12285866
申请日:2008-10-15
申请人: Kiyozo Asada , Takashi Uemori , Yoshimi Sato , Tomoko Fujita , Kazue Miyake , Osamu Takeda , Hiroyuki Mukai , Ikunoshin Kato
发明人: Kiyozo Asada , Takashi Uemori , Yoshimi Sato , Tomoko Fujita , Kazue Miyake , Osamu Takeda , Hiroyuki Mukai , Ikunoshin Kato
CPC分类号: C12P19/34 , C12N9/1252 , C12N9/96 , C12Q1/6806 , Y10S435/975 , C12Q2521/107
摘要: A DNA synthesis reaction-enhancer comprising at least one kind selected from the group consisting of acidic substances and cationic complexes; a DNA synthesis method in which during a DNA synthesis reaction a reaction is carried out in the presence of the above enhancer by using DNA polymerase; a DNA synthesis reaction composition comprising the above enhancer; a DNA synthesis reaction composition comprising two or more kinds of DNA polymerases each having 3′→5′ exonuclease activity; a DNA synthesis method in which during a DNA synthesis reaction two or more kinds of DNA polymerases each having 3′→5′ exonuclease activity are used; a kit for use in in vitro DNA synthesis, comprising two or more kinds of DNA polymerases each having 3′→5′ exonuclease activity; and a kit for use in in vitro DNA synthesis, wherein the kit comprises the DNA synthesis reaction-enhancer and DNA polymerase. According to the present invention, DNA synthesis can be carried out at an efficiency more excellent as compared to conventional DNA synthesis reaction.
摘要翻译: 一种DNA合成反应增强剂,其包含选自酸性物质和阳离子配合物中的至少一种; DNA合成方法,其中在DNA合成反应中,通过使用DNA聚合酶在上述增强剂的存在下进行反应; 包含上述增强剂的DNA合成反应组合物; 包含两种以上各自具有3'→5'核酸外切酶活性的DNA聚合酶的DNA合成反应组合物; 一种DNA合成方法,其中在DNA合成反应中使用两种或更多种具有3'→5'核酸外切酶活性的DNA聚合酶; 用于体外DNA合成的试剂盒,其包含两种或更多种各自具有3'→5'核酸外切酶活性的DNA聚合酶; 以及用于体外DNA合成的试剂盒,其中试剂盒包含DNA合成反应增强剂和DNA聚合酶。 根据本发明,与常规DNA合成反应相比,可以以更优异的效果进行DNA合成。
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公开(公告)号:US20070298415A1
公开(公告)日:2007-12-27
申请号:US10582345
申请日:2004-12-06
申请人: Takashi Uemori , Osamu Takeda , Junko Yamamoto , Hiroyuki Mukai , Kiyozo Asada , Ikunoshin Kato
发明人: Takashi Uemori , Osamu Takeda , Junko Yamamoto , Hiroyuki Mukai , Kiyozo Asada , Ikunoshin Kato
CPC分类号: C12Q1/6844 , C12Q2531/119 , C12Q2525/185 , C12Q2537/143 , C12Q2525/161
摘要: In the case where a template nucleic acid has sequences substantially the same as each other, a primer is designed in a region between these sequences so as to give a product in a ladder structure in which a target region is polymerized in a single sequence via the same sequences in the ICAN reaction. By using a chimeric oligonucleotide primer and a ladder-forming oligonucleotide primer containing a specific base sequence, an amplification product having a ladder structure can be positively formed and thus the sensitivity, amplification efficiency and reaction speed in the ICAN reaction can be elevated.
摘要翻译: 在模板核酸具有彼此基本相同的序列的情况下,在这些序列之间的区域中设计引物,以便产生梯形结构中的产物,其中靶区域通过经由 在ICAN反应中相同的序列。 通过使用嵌合寡核苷酸引物和含有特定碱基序列的梯形寡核苷酸引物,可以积极地形成具有梯形结构的扩增产物,从而可以提高ICAN反应中的灵敏度,扩增效率和反应速度。
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公开(公告)号:US20050118578A1
公开(公告)日:2005-06-02
申请号:US10467325
申请日:2002-02-06
申请人: Junichi Mineno , Osamu Takeda , Masatomo Rokushima , Takashi Uemori , Shigekazu Hokazono , Hiroyuki Mukai , Shusaku Yamashita , Hiroaki Sagawa , Kiyozo Asada , Ikunoshin Kato
发明人: Junichi Mineno , Osamu Takeda , Masatomo Rokushima , Takashi Uemori , Shigekazu Hokazono , Hiroyuki Mukai , Shusaku Yamashita , Hiroaki Sagawa , Kiyozo Asada , Ikunoshin Kato
CPC分类号: C12Q1/6837 , C12Q2565/518 , C12Q2531/119
摘要: A nucleic acid, which is provided in a large amount through a nucleic acid amplification reaction with the use of chimeric oligonucleotide primers, is constructed in a state of containing a modified deoxyribonucleotide for immobilizing the nucleic acid to a solid phase and then immobilized to a solid phase at a high efficiency, thereby giving an immobilized nucleic acid product with excellent qualities.
摘要翻译: 通过使用嵌合寡核苷酸引物通过核酸扩增反应大量提供的核酸以含有修饰的脱氧核糖核苷酸的状态构建,用于将核酸固定至固相,然后固定于固体 相,从而得到具有优异品质的固定化核酸产物。
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公开(公告)号:US06218150B1
公开(公告)日:2001-04-17
申请号:US09446504
申请日:1999-12-23
申请人: Takashi Uemori , Yoshimi Sato , Tomoko Fujita , Kazue Miyake , Hiroyuki Mukai , Kiyozo Asada , Ikunoshin Kato
发明人: Takashi Uemori , Yoshimi Sato , Tomoko Fujita , Kazue Miyake , Hiroyuki Mukai , Kiyozo Asada , Ikunoshin Kato
IPC分类号: C12P1934
CPC分类号: C12N9/1252
摘要: The present invention relates to a thermostable DNA polymerase-associated factor capable of enhancing DNA synthesizing-activity of a DNA polymerase; a thermostable DNA polymerase-associated factor possessing an activity of binding to a DNA polymerase and a method for producing the same; a gene encoding the DNA polymerase-associated factor; a method of DNA synthesis by using a DNA polymerase in the presence of the DNA polymerase-associated factor; and a kit comprising the DNA polymerase-associated factor. According to the present invention, there can be provided in vitro DNA synthesis and a DNA amplification system which are more excellent than conventional techniques by utilizing the DNA polymerase-associated factor of the present invention.
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公开(公告)号:US07521178B1
公开(公告)日:2009-04-21
申请号:US09673884
申请日:1999-04-21
申请人: Kiyozo Asada , Takashi Uemori , Yoshimi Sato , Tomoko Fujita , Kazue Miyake , Osamu Takeda , Hiroyuki Mukai , Ikunoshin Kato
发明人: Kiyozo Asada , Takashi Uemori , Yoshimi Sato , Tomoko Fujita , Kazue Miyake , Osamu Takeda , Hiroyuki Mukai , Ikunoshin Kato
CPC分类号: C12P19/34 , C12N9/1252 , C12N9/96 , C12Q1/6806 , Y10S435/975 , C12Q2521/107
摘要: A DNA synthesis reaction-enhancer comprising at least one kind selected from the group consisting of acidic substances and cationic complexes; a DNA synthesis method in which during a DNA synthesis reaction a reaction is carried out in the presence of the above enhancer by using DNA polymerase; a DNA synthesis reaction composition comprising the above enhancer; a DNA synthesis reaction composition comprising two or more kinds of DNA polymerases each having 3′→5′ exonuclease activity; a DNA synthesis method in which during a DNA synthesis reaction two or more kinds of DNA polymerases each having 3′→5′ exonuclease activity are used; a kit for use in in vitro DNA synthesis, comprising two or more kinds of DNA polymerases each having 3′→5′ exonuclease activity; and a kit for use in in vitro DNA synthesis, wherein the kit comprises the DNA synthesis reaction-enhancer and DNA polymerase. According to the present invention, DNA synthesis can be carried out at an efficiency more excellent as compared to conventional DNA synthesis reaction.
摘要翻译: 一种DNA合成反应增强剂,其包含选自酸性物质和阳离子配合物中的至少一种; DNA合成方法,其中在DNA合成反应中,通过使用DNA聚合酶在上述增强剂的存在下进行反应; 包含上述增强剂的DNA合成反应组合物; 包含两种或更多种各自具有3'→5'核酸外切酶活性的DNA聚合酶的DNA合成反应组合物; 一种DNA合成方法,其中在DNA合成反应中使用两种或更多种具有3'→5'核酸外切酶活性的DNA聚合酶; 用于体外DNA合成的试剂盒,其包含两种或更多种各自具有3'→5'核酸外切酶活性的DNA聚合酶; 和用于体外DNA合成的试剂盒,其中试剂盒包含DNA合成反应增强剂和DNA聚合酶。 根据本发明,与常规DNA合成反应相比,可以以更优异的效果进行DNA合成。
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公开(公告)号:US06673578B1
公开(公告)日:2004-01-06
申请号:US09786684
申请日:2001-05-22
申请人: Takashi Uemori , Yoshimi Sato , Mariko Okawa , Tomoko Fujita , Kazue Miyake , Osamu Takeda , Hiroaki Sagawa , Michio Hagiya , Hiroyuki Mukai , Kiyozo Asada , Ikunoshin Kato
发明人: Takashi Uemori , Yoshimi Sato , Mariko Okawa , Tomoko Fujita , Kazue Miyake , Osamu Takeda , Hiroaki Sagawa , Michio Hagiya , Hiroyuki Mukai , Kiyozo Asada , Ikunoshin Kato
IPC分类号: C12P1934
CPC分类号: C12N15/1003 , C12Q1/686 , C12Q2527/125
摘要: A DNA synthesis method with a shortened time period required for DNA synthesis by polymerase chain reaction (PCR), characterized in that a DNA polymerase is used in an amount effective for providing more than 10 ng of amplified DNA fragments of about 2 kb per 50 &mgr;l of a reaction mixture, when PCR is carried out under the following conditions (A) and (B): (A) reaction mixture: 50 &mgr;l volume of a reaction mixture comprising DNA polymerase, 1 ng of genomic DNA from Escherichia coli, and 10 pmol each of primers Eco-1 and Eco-2 (nucleotide sequences of the primers Eco-1 and Eco-2 being shown in SEQ ID NOs: 10 and 11 of Sequence Listing, respectively); and having a composition suitable for the DNA polymerase; and (B) reaction conditions: 35 cycles of PCR, wherein one cycle consists of 99° C., 1 second-66° C., 7 seconds; a kit for DNA synthesis usable for the DNA synthesis method; and an article of manufacture of a PCR agent. According to the present invention, the procedures in the genetic engineering studies and industries involved with PCR can be speeded up.
摘要翻译: 一种DNA合成方法,其通过聚合酶链式反应(PCR)进行DNA合成所需的时间缩短,其特征在于使用DNA聚合酶,其量有效提供超过10ng每50ul约2kb的扩增DNA片段 的反应混合物,当在以下条件(A)和(B)下进行PCR时:(A)反应混合物:将50体积的包含DNA聚合酶的反应混合物,1ng来自大肠杆菌的基因组DNA和10ng 各引物Eco-1和Eco-2(引物Eco-1和Eco-2的核苷酸序列分别示于序列表的SEQ ID NO:10和11中) 并具有适合于DNA聚合酶的组合物; 和(B)反应条件:35个循环的PCR,其中一个循环由99℃,1秒-66℃,7秒; 用于DNA合成方法的DNA合成试剂盒; 以及PCR试剂的制造。 根据本发明,可以加快基因工程研究和涉及PCR的行业的程序。
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公开(公告)号:US5366883A
公开(公告)日:1994-11-22
申请号:US894212
申请日:1992-06-09
申请人: Kiyozo Asada , Takashi Uemori , Hiroyuki Mukai , Ikunoshin Kato , Kenneth Laderman , Christian B. Anfinsen
发明人: Kiyozo Asada , Takashi Uemori , Hiroyuki Mukai , Ikunoshin Kato , Kenneth Laderman , Christian B. Anfinsen
IPC分类号: C07K14/195 , C12N1/21 , C12N9/26 , C12N9/28 , C12N15/09 , C12N15/56 , C12R1/125 , C12R1/19 , C12N15/63 , C12N15/75
CPC分类号: C12N9/2417 , C07K14/195
摘要: The present invention relates, in general, to a cloned .alpha.-amylase gene, and, in particular, to a cloned hyperthermophilic .alpha.-amylase gene and to methods of producing .alpha.-amylase using same.
摘要翻译: 本发明一般涉及克隆的α-淀粉酶基因,特别是涉及克隆的超嗜热α-淀粉酶基因以及使用其生产α-淀粉酶的方法。
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