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1.发明公开
公开(公告)号：EP1111044A1
公开(公告)日：2001-06-27
申请号：EP99940673.9
申请日：1999-09-06
发明人： UEMORI, Takashi , SATO, Yoshimi , OKAWA, Mariko , FUJITA, Tomoko , MIYAKE, Kazue , TAKEDA, Osamu , SAGAWA, Hiroaki , HAGIYA, Michio , MUKAI, Hiroyuki , ASADA, Kiyozo , KATO, Ikunoshin
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 µl of a reaction mixture, when PCR is carried out under the following conditions (A) and (B): (A) reaction mixture: 50 µl 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： 一种通过聚合酶链式反应（PCR）进行DNA合成所需的时间缩短的DNA合成方法，其特征在于使用DNA聚合酶，其用量可提供超过10ng的扩增DNA片段，每50ul约2kb （A）和（B）：（A）反应混合物：50μl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）的10pmol; 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; 和PCR试剂的制造。 根据本发明，可以加快基因工程研究中涉及PCR的工艺步骤。
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公开(公告)号：EP1072678A1
公开(公告)日：2001-01-31
申请号：EP99917081.4
申请日：1999-04-21
发明人： ASADA, Kiyozo , UEMORI, Takashi , SATO, Yoshimi , FUJITA, Tomoko , MIYAKE, Kazue , TAKEDA, Osamu , MUKAI, Hiroyuki , KATO, Ikunoshin
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聚合酶在上述增强子的存在下进行反应; 包含上述增强剂的DNA合成反应组合物; 一种DNA合成反应组合物，其包含两种或更多种各自具有3个→5个核酸外切酶活性的DNA聚合酶; 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个→5个核酸外切酶活性的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个→5个核酸外切酶活性的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：EP0997530A1
公开(公告)日：2000-05-03
申请号：EP98929687.6
申请日：1998-06-24
发明人： UEMORI, Takashi , SATO, Yoshimi , FUJITA, Tomoko , MIYAKE, Kazue , MUKAI, Hiroyuki , ASADA, Kiyozo , KATO, Ikunoshin
IPC分类号： C12N15/54 , C12N9/12 , C12N15/31 , C07K14/195 , C12P21/02
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.
摘要翻译： 本发明涉及能够增强DNA聚合酶的DNA合成活性的耐热性DNA聚合酶相关因子; 具有与DNA聚合酶结合的活性的热稳定DNA聚合酶相关因子及其制备方法; 编码DNA聚合酶相关因子的基因; 在DNA聚合酶相关因子存在下使用DNA聚合酶进行DNA合成的方法; 和包含DNA聚合酶相关因子的试剂盒。 根据本发明，通过利用本发明的DNA聚合酶相关因子，可以提供比常规技术更优异的体外DNA合成和DNA扩增系统。
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