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公开(公告)号：US20120164654A1
公开(公告)日：2012-06-28
申请号：US13393302
申请日：2010-08-27
申请人： Yuko Nakabayashi , Yoshimi Sato , Takashi Uemori , Hiroyuki Mukai
发明人： Yuko Nakabayashi , Yoshimi Sato , Takashi Uemori , Hiroyuki Mukai
CPC分类号： C12Q1/686 , C12N15/1096 , C12Q2527/125 , C12Q2521/107
摘要： A composition for a reverse transcription polymerase chain reaction, which comprises a thermostable DNA polymerase, a reverse transcriptase, a dye marker and a specific gravity-increasing agent; and a premix reagent for a one-step RT-PCR, which comprises the composition, is not frozen under usual storage conditions at −20 to −30° C. and has excellent handleability.
摘要翻译： 用于逆转录聚合酶链反应的组合物，其包含热稳定性DNA聚合酶，逆转录酶，染料标记物和比重增加剂; 并且包含该组合物的用于一步RT-PCR的预混合试剂在-20至-30℃的通常储存条件下不冷冻，并且具有优异的可操作性。
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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：US08344105B2
公开(公告)日：2013-01-01
申请号：US13239981
申请日：2011-09-22
申请人： Yoshimi Sato , Kazue Nishiwaki , Nana Shimada , Shigekazu Hokazono , Takashi Uemori , Hiroyuki Mukai , Ikunoshin Kato
发明人： Yoshimi Sato , Kazue Nishiwaki , Nana Shimada , Shigekazu Hokazono , Takashi Uemori , Hiroyuki Mukai , Ikunoshin Kato
CPC分类号： C12Q1/686 , C12N9/1252 , C12Q2521/101
摘要： A polypeptide having a high fidelity DNA polymerase activity and thus being useful as a genetic engineering reagent; a gene encoding this polypeptide; a method of producing the polypeptide; and a method of amplifying a nucleic acid by using the polypeptide.
摘要翻译： 具有高保真DNA聚合酶活性的多肽，因此可用作遗传工程试剂; 编码该多肽的基因; 一种产生该多肽的方法; 以及通过使用多肽扩增核酸的方法。
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公开(公告)号：US08048987B2
公开(公告)日：2011-11-01
申请号：US12708111
申请日：2010-02-18
申请人： Yoshimi Sato , Kazue Nishiwaki , Nana Shimada , Shigekazu Hokazono , Takashi Uemori , Hiroyuki Mukai , Ikunoshin Kato
发明人： Yoshimi Sato , Kazue Nishiwaki , Nana Shimada , Shigekazu Hokazono , Takashi Uemori , Hiroyuki Mukai , Ikunoshin Kato
CPC分类号： C12Q1/686 , C12N9/1252 , C12Q2521/101
摘要： A polypeptide having a high fidelity DNA polymerase activity and thus being useful as a genetic engineering reagent; a gene encoding this polypeptide; a method of producing the polypeptide; and a method of amplifying a nucleic acid by using the polypeptide.
摘要翻译： 具有高保真DNA聚合酶活性的多肽，因此可用作遗传工程试剂; 编码该多肽的基因; 一种产生该多肽的方法; 以及通过使用多肽扩增核酸的方法。
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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：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：US07846703B2
公开(公告)日：2010-12-07
申请号：US11866148
申请日：2007-10-02
申请人： Eiji Kobayashi , Yuki Ueda , Yoshimi Sato , Takashi Uemori , Hiroyuki Mukai , Ikunoshin Kato
发明人： Eiji Kobayashi , Yuki Ueda , Yoshimi Sato , Takashi Uemori , Hiroyuki Mukai , Ikunoshin Kato
CPC分类号： C12N9/1241
摘要： A polymerase activity is effectively enhanced by adding an anionic surfactant, in particular an anionic surfactant having a polyethoxyl group, to a reaction mixture containing a polymerase.
摘要翻译： 通过向含有聚合酶的反应混合物中加入阴离子表面活性剂，特别是具有聚乙氧基的阴离子表面活性剂，可有效提高聚合酶活性。
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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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