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    Detection Method of Dna Amplification Using Probe Labeled With Intercalating Dye
    1.
    发明申请
    Detection Method of Dna Amplification Using Probe Labeled With Intercalating Dye 审中-公开
    用插入染料标记的探针的Dna扩增检测方法

    公开(公告)号：US20080220415A1

    公开(公告)日：2008-09-11

    申请号：US10593900

    申请日：2005-03-25

    申请人： Han Oh Park ,  Hyun-Bae Kim ,  Sung-Min Chi

    发明人： Han Oh Park ,  Hyun-Bae Kim ,  Sung-Min Chi

    IPC分类号： C12Q1/68

    CPC分类号： C12Q1/6844 ,  C12Q1/686 ,  C12Q2563/173 ,  C12Q2563/107 ,  C12Q2561/113

    摘要： The present invention relates to a detection method of nucleic acid amplification using probe labeled with intercalating dye. More particularly, the present invention is directed to a real-time detection method of nucleic acid amplification, comprising the steps of i) producing an aqueous buffer which contains a nucleic acid, a pair of primers for amplification of said nucleic acid, a fluorescent probe wherein a fluorescent dye of which intensity of fluorescence is varied when the dye is intercalated into a double-stranded nucleic acid, is connected with an oligonucleotide of which base sequence is complementary with at least a part of said nucleic acid, four (4) kinds of nucleotides and DNA polymerase; ii) denaturing said doublestranded nucleic acid into single strands by heating the aqueous buffer prepared in step i) up to 931 C to 96 C; iii) annealing said pair of primers with said single strand by cooling the solution obtained in step ii) up to 50 C. to 571 C; iv) replicating said single-stranded nucleic acid by heating the solution obtained from step iii) up to 701 C to 74° C.; v) denaturing said replicated nucleic add into single strands by heating the solution obtained in step iv) up to 931 C to 961 C; vi) annealing said fluorescent probe with said single-stranded nucleic acid by cooling the solution obtained in step v up to 501 C to 57 C; vii) measuring an intensity of the fluorescence emitted from the solution obtained in step vi); and viii) repeating more than one steps iv) through vii).

    摘要翻译： 本发明涉及使用用插层染料标记的探针的核酸扩增检测方法。 更具体地，本发明涉及核酸扩增的实时检测方法，包括以下步骤：i）产生含有核酸的水性缓冲液，用于扩增所述核酸的一对引物，荧光探针 其中当染料插入双链核酸时荧光强度变化的荧光染料与其碱基序列与所述核酸的至少一部分互补的寡核苷酸连接，四（4）种 的核苷酸和DNA聚合酶; ii）通过加热步骤i）中制备的含水缓冲液至931℃至96℃，将所述双链核酸变性为单链; iii）通过将步骤ii）中获得的溶液冷却至50℃至571℃，用所述单链退火所述一对引物; iv）通过将从步骤iii）获得的溶液加热至701℃至74℃来复制所述单链核酸; v）通过将步骤iv）中获得的溶液加热至931℃至961℃，将所述复制的核酸添加到单链中; vi）通过将步骤v中获得的溶液冷却至501℃至57℃，用所述单链核酸退火所述荧光探针; vii）测量从步骤vi）中获得的溶液发射的荧光的强度; 和viii）重复多于一个步骤iv）至vii）。

    Primers for PCR amplification comprising a basic parts within the primer sequences
    2.
    发明授权
    Primers for PCR amplification comprising a basic parts within the primer sequences 有权
    用于PCR扩增的引物，其包含引物序列内的基本部分

    公开(公告)号：US08513399B2

    公开(公告)日：2013-08-20

    申请号：US12681754

    申请日：2008-10-02

    申请人： Hyun Bae Kim ,  Seong Youl Kim ,  Jun Mo Gil ,  Hae Joon Park ,  Han Oh Park

    发明人： Hyun Bae Kim ,  Seong Youl Kim ,  Jun Mo Gil ,  Hae Joon Park ,  Han Oh Park

    IPC分类号： C07H21/04 ,  C07H19/04 ,  C12Q1/68 ,  C12P19/34

    CPC分类号： C12Q1/686 ,  C12Q2525/119

    摘要： The present invention relates to primers for PCR amplification comprising abasic parts within the primer sequences and a method for PCR amplification using the same. More precisely, the present invention relates to primers capable of amplifying different templates and having abasic parts complementary to mutated site or polymorphic site of template DNA and a method for PCR amplification comprising the steps of mixing the composition for PCR amplification comprising the primers with nucleic acid template; and performing PCR with the mixture. The primers for PCR amplification of the present invention contain abasic parts not having specific coding information in their nucleotide sequences, so that they can amplify different templates having mutated sites at the same time.

    摘要翻译： 本发明涉及用于PCR扩增的引物，其包括引物序列内的无碱部分和使用其的PCR扩增方法。 更确切地说，本发明涉及能够扩增不同模板并具有与模板DNA的突变位点或多态位点互补的脱碱基的引物，以及用于PCR扩增的方法，其包括将包含引物的PCR扩增组合物与核酸 模板; 并用混合物进行PCR。 用于本发明的PCR扩增的引物含有在其核苷酸序列中不具有特定编码信息的脱碱基，从而可以同时扩增具有突变位点的不同模板。

    DRIED COMPOSITION FOR HOT-START PCR WITH LONG-TERM STABILITY
    3.
    发明申请
    DRIED COMPOSITION FOR HOT-START PCR WITH LONG-TERM STABILITY 有权
    用于具有长期稳定性的热启动PCR的干燥组合物

    公开(公告)号：US20100209973A1

    公开(公告)日：2010-08-19

    申请号：US12682456

    申请日：2008-10-28

    申请人： Seong-Youl Kim ,  Hyun Bae Kim ,  Hae-Joon Park ,  Han Oh Park

    发明人： Seong-Youl Kim ,  Hyun Bae Kim ,  Hae-Joon Park ,  Han Oh Park

    IPC分类号： C12P19/34 ,  C12N9/14

    CPC分类号： C12Q1/686 ,  C12Q2549/101 ,  C12Q2527/125

    摘要： The present invention relates to a dried composition for hot-start PCR, more precisely a dried composition for hot-start PCR with improved stability and long-term storagability which is characteristically prepared by the steps of preparing a reaction mixture by mixing an aqueous solution containing reaction buffer, MgCl2, 4 types of dNTPs, DNA polymerase with pyrophosphate and pyrophosphatase in a reaction tube; and drying the reaction mixture prepared above, a preparation method of the same and a method for amplifying nucleic acid using the same. The dried composition for hot-start PCR is added with pyrophosphate and pyrophosphatase together before drying, so that it can have improved stability and long-term storagability as well as convenience in use, compared with the conventional compositions for hot-start PCR. Therefore, this composition can be effectively used for hot-start PCR, multiplex PCR or real-time quantitative PCR.

    摘要翻译： 本发明涉及一种用于热启动PCR的干燥组合物，更准确地说是一种用于热启动PCR的干燥组合物，其具有改进的稳定性和长期储存性，其特征在于通过以下步骤制备反应混合物：将含有 反应缓冲液，MgCl 2，4种dNTP，反应管中含有焦磷酸的DNA聚合酶和焦磷酸酶; 并干燥上述制备的反应混合物，其制备方法和使用其制备用于扩增核酸的方法。 用于热启动PCR的干燥组合物在干燥之前加入焦磷酸盐和焦磷酸酶，与常规的热启动组合物相比，其可以具有改善的稳定性和长期储存性以及使用方便性。 因此，该组合物可有效用于热启动PCR，多重PCR或实时定量PCR。

    Dried composition for hot-start PCR with long-term stability
    4.
    发明授权
    Dried composition for hot-start PCR with long-term stability 有权
    用于热启动PCR的干组合物具有长期稳定性

    公开(公告)号：US09034603B2

    公开(公告)日：2015-05-19

    申请号：US12682456

    申请日：2008-10-28

    申请人： Seong-Youl Kim ,  Hyun Bae Kim ,  Hae-Joon Park ,  Han Oh Park

    发明人： Seong-Youl Kim ,  Hyun Bae Kim ,  Hae-Joon Park ,  Han Oh Park

    IPC分类号： C12Q1/68

    CPC分类号： C12Q1/686 ,  C12Q2549/101 ,  C12Q2527/125

    摘要： The present invention relates to a dried composition for hot-start PCR, more precisely a dried composition for hot-start PCR with improved stability and long-term storagability which is characteristically prepared by the steps of preparing a reaction mixture by mixing an aqueous solution containing reaction buffer, MgCl2, 4 types of dNTPs, DNA polymerase with pyrophosphate and pyrophosphatase in a reaction tube; and drying the reaction mixture prepared above, a preparation method of the same and a method for amplifying nucleic acid using the same. The dried composition for hot-start PCR is added with pyrophosphate and pyrophosphatase together before drying, so that it can have improved stability and long-term storagability as well as convenience in use, compared with the conventional compositions for hot-start PCR. Therefore, this composition can be effectively used for hot-start PCR, multiplex PCR or real-time quantitative PCR.

    摘要翻译： 本发明涉及一种用于热启动PCR的干燥组合物，更准确地说是一种用于热启动PCR的干燥组合物，其具有改进的稳定性和长期储存性，其特征在于通过以下步骤制备反应混合物：将含有 反应缓冲液，MgCl 2，4种dNTP，反应管中含有焦磷酸的DNA聚合酶和焦磷酸酶; 并干燥上述制备的反应混合物，其制备方法和使用其制备用于扩增核酸的方法。 用于热启动PCR的干燥组合物在干燥之前加入焦磷酸盐和焦磷酸酶，与常规的热启动组合物相比，其可以具有改善的稳定性和长期储存性以及使用方便性。 因此，该组合物可有效用于热启动PCR，多重PCR或实时定量PCR。

    PRIMERS FOR PCR AMPLIFICATION COMPRISING A BASIC PARTS WITHIN THE PRIMER SEQUENCES
    5.
    发明申请
    PRIMERS FOR PCR AMPLIFICATION COMPRISING A BASIC PARTS WITHIN THE PRIMER SEQUENCES 有权
    用于在扩展序列中包含基本部分的PCR扩增的引物

    公开(公告)号：US20110008845A1

    公开(公告)日：2011-01-13

    申请号：US12681754

    申请日：2008-10-02

    申请人： Hyun Bae Kim ,  Seong Youl Kim ,  Jun Mo Gil ,  Hae Joon Park ,  Han Oh Park

    发明人： Hyun Bae Kim ,  Seong Youl Kim ,  Jun Mo Gil ,  Hae Joon Park ,  Han Oh Park

    IPC分类号： C12P19/34 ,  C07H21/00 ,  C12N9/12

    CPC分类号： C12Q1/686 ,  C12Q2525/119

    摘要： The present invention relates to primers for PCR amplification comprising abasic parts within the primer sequences and a method for PCR amplification using the same. More precisely, the present invention relates to primers capable of amplifying different templates and having abasic parts complementary to mutated site or polymorphic site of template DNA and a method for PCR amplification comprising the steps of mixing the composition for PCR amplification comprising the primers with nucleic acid template; and performing PCR with the mixture. The primers for PCR amplification of the present invention contain abasic parts not having specific coding information in their nucleotide sequences, so that they can amplify different templates having mutated sites at the same time.

    摘要翻译： 本发明涉及用于PCR扩增的引物，其包括引物序列内的无碱部分和使用其的PCR扩增方法。 更确切地说，本发明涉及能够扩增不同模板并具有与模板DNA的突变位点或多态位点互补的脱碱基的引物，以及用于PCR扩增的方法，其包括将包含引物的PCR扩增组合物与核酸 模板; 并用混合物进行PCR。 用于本发明的PCR扩增的引物含有在其核苷酸序列中不具有特定编码信息的脱碱基，从而可以同时扩增具有突变位点的不同模板。

    Automatic necleic acid purification apparatus and method for aerosol-protecting
    6.
    发明授权
    Automatic necleic acid purification apparatus and method for aerosol-protecting 有权
    自动碳酸净化装置及气溶胶保护方法

    公开(公告)号：US09273306B2

    公开(公告)日：2016-03-01

    申请号：US13885556

    申请日：2011-11-18

    申请人： Han Oh Park ,  Byung Rae Jeong ,  Kwon Sic Kim

    发明人： Han Oh Park ,  Byung Rae Jeong ,  Kwon Sic Kim

    IPC分类号： C12Q1/68 ,  B01L3/00 ,  C12N15/10

    CPC分类号： C12N15/1013 ,  C12Q1/68

    摘要： Disclosed is an automatic nucleic acid purification apparatus which can prevent pollution due to aerosol generated from a biological sample containing high concentration target nucleic acid when the biological sample containing the high concentration target nucleic acid is mixed with other biological sample containing low concentration target nucleic acid or not containing the target nucleic acid. Further, disclosed is an automatic nucleic acid purification apparatus which can be applied to all kinds of nucleic acid purification equipments for purifying a plurality of biological samples using a magnet rode or a multi-pipette block moving in two or three axial directions, and which can minimize pollution due to the aerosol generated from the biological sample containing high concentration target nucleic acid and also can obtain accurate results.

    摘要翻译： 公开了一种自动核酸纯化装置，当含有高浓度靶核酸的生物样品与含有低浓度靶核酸的其他生物样品混合时，可以防止由含有高浓度靶核酸的生物样品产生的气溶胶引起的污染，或 不含靶核酸。 此外，公开了一种自动核酸纯化装置，其可以应用于使用磁珠或多个移液管块在两个或三个轴向移动的多个生物样品的各种核酸纯化设备，并且其可以 尽可能减少由含有高浓度靶核酸的生物样品产生的气溶胶引起的污染，并可获得准确的结果。

    METHOD OF IDENTIFYING NUCLEIC ACID-CONTAINING OBJECT
    9.
    发明申请
    METHOD OF IDENTIFYING NUCLEIC ACID-CONTAINING OBJECT 审中-公开
    识别含有核酸的物体的方法

    公开(公告)号：US20140087377A1

    公开(公告)日：2014-03-27

    申请号：US14005213

    申请日：2012-03-13

    申请人： Han Oh Park ,  Jung Young Choi ,  Eun Su Han ,  Gu Young Song ,  Won Seok Jang

    发明人： Han Oh Park ,  Jung Young Choi ,  Eun Su Han ,  Gu Young Song ,  Won Seok Jang

    IPC分类号： C12Q1/68

    CPC分类号： C12Q1/6816 ,  C12Q1/6818 ,  C12Q2521/327 ,  C12Q2525/121 ,  C12Q2563/185

    摘要： The present invention relates to a method of identifying nucleic acid-containing object, more precisely a method of identifying nucleic acid-containing object which comprises the following steps: (1) preparing nucleic acid-containing object having the nucleotide sequence complementary to the nucleotide sequence of RNA-dual probe; (2) reacting the nucleic acid included in the object with the buffer containing the RNA-dual probe conjugated with a reporter and a quencher respectively and RNase; and (3) detecting fluorescence generated from the reporter. The method of identifying an object of the present invention provides labeling sensitivity 100 times as high as that of the conventional method using sequencing or labeling with fluorescent materials, takes advantages of shorter analysis time, facilitates different labeling on a variety of products according to fluorescent materials, and makes possible unlimited product administration by product group and batch in real production process by differentiating the nucleotide sequence of each oligonucleotide.

    摘要翻译： 本发明涉及鉴定含核酸物质的方法，更准确地说，一种鉴定含核酸物质的方法，包括以下步骤：（1）制备具有与核苷酸序列互补的核苷酸序列的含核酸物质 的RNA双探针; （2）将包含在物体中的核酸与分别与报告子和猝灭剂结合的RNA双重探针的缓冲液和RNase分别反应; 和（3）检测从报告物产生的荧光。 识别本发明的目的的方法提供了使用荧光材料进行测序或标记的常规方法的100倍的标签灵敏度，具有较短的分析时间的优点，便于根据荧光材料对各种产品进行不同的标记 并通过区分每个寡核苷酸的核苷酸序列，在实际生产过程中使产品组和批次无限制地进行产品管理。