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    Method for synthesizing DNA
    5.
    发明授权
    Method for synthesizing DNA 失效
    DNA合成方法

    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。

    Method of Amplifying Nucleic Acid
    7.
    发明申请
    Method of Amplifying Nucleic Acid 审中-公开
    核酸扩增方法

    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

    IPC分类号： C12P19/34 ,  C07H21/04 ,  C12Q1/68

    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。

    Method for synthesizing DNA
    8.
    发明授权
    Method for synthesizing DNA 有权
    DNA合成方法

    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

    IPC分类号： C12Q1/68 ,  C12P19/34 ,  C07H21/02 ,  C07H21/04

    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。

    Method for synthesizing DNA
    10.
    发明授权
    Method for synthesizing DNA 有权
    DNA合成方法

    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

    IPC分类号： C12Q1/68 ,  C12P19/34 ,  C07H21/02

    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。