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    • 3. 发明公开
    • 어시스트프로브 및 그의 이용방법
    • 辅助探讨AMD使用方法
    • KR1020070106705A
    • 2007-11-05
    • KR1020077017804
    • 2006-02-27
    • 에자이 알앤드디 매니지먼트 가부시키가이샤
    • 후지카와토시히코우스이,미츠구
    • C12N15/09C12Q1/68G01N33/566
    • C12N15/10C12N15/1068C12Q1/682C12Q1/6876C12Q2521/107C12Q2525/161C12Q2525/173C12Q2533/107C12Q2537/125C12Q2537/143C12Q2537/162
    • It is intended to provide a method of detecting target genes whereby the sensitivity in the pulser method can be improved and multiple genes can be simultaneously detected, assist probes to be used in the above method, and a method of forming a signal probe polymer by using the above assist probes. A method of detecting target genes by forming a signal probe polymer with the use of a first probe which has a nucleic acid region X, a nucleic acid region Y and a nucleic acid region Z in this order from the 5' end, a second probe which has a nucleic acid region X', a nucleic acid region Y' and a nucleic acid region Z' in this order from the 5' end and a plural number of assist probes which have target regions being hybridizable with the same nucleic acid region and target gene as in the first probe as described above. In this method, the assist probes are designed so as to have a structure comprising the above-described nucleic acid regions X, Y and X and the above-described target region in this order from the 5' end or a structure comprising the above-described target region and the above-described nucleic acid regions Z, Y and Z in this order from the 5' end.
    • 旨在提供一种检测靶基因的方法,由此能够改善脉冲发生器方法的灵敏度并可以同时检测多个基因,在上述方法中使用辅助探针,以及通过使用以下方法形成信号探针聚合物的方法: 上述辅助探针。 通过使用第一探针形成信号探针聚合物来检测靶基因的方法,所述第一探针从5'端开始依次具有核酸区域X,核酸区域Y和核酸区域Z,第二探针 其具有从5'端起依次具有核酸区域X',核酸区域Y'和核酸区域Z'的多个辅助探针和具有可与相同核酸区域杂交的靶区域的多个辅助探针,以及 靶基因如上所述的第一探针中。 在该方法中,辅助探针被设计为具有包含上述核酸区域X,Y和X以及上述目标区域的结构,从5'端开始依次包括上述靶区域, 上述目标区域和上述核酸区域Z,Y和Z从5'端开始。
    • 4. 发明授权
    • 단일 뉴클레오티드 검출 방법
    • 单核苷酸检测方法
    • KR101754091B1
    • 2017-07-05
    • KR1020177001507
    • 2015-07-22
    • 베이스4 이노베이션 엘티디
    • 밤포스,바너비프레일링,카메론알렉산더
    • C12Q1/68
    • C12Q1/6869C12Q1/6823C12Q2521/101C12Q2521/319C12Q2521/501C12Q2525/125C12Q2525/307C12Q2533/107C12Q2537/149C12Q2537/162C12Q2563/107C12Q2563/159C12Q2565/629
    • DNA 또는 RNA와같은핵산의염기서열분석방법을제공한다. 본실시예는 (1) 상기핵산의점진적인피로인산분해에의해단일뉴클레오시드 3인산스트림을생성하는단계; (2) 중합효소및 리가아제의존재하에적어도하나의상기단일뉴클레오시드 3인산과상응하는프로브시스템을반응시킴으로써적어도하나의실질적인이중가닥올리고뉴클레오티드사용프로브(used probe)를생성하는단계로, 상기상응하는프로브시스템은, (a) 검출불가능한상태의특이적검출가능요소들로표지된단일가닥제1 올리고뉴클레오티드및 (b) 상기제1 올리고뉴클레오티드상의상보적영역에혼성화할수 있는단일가닥제2 및제3 올리고뉴클레오티드를포함하는단계; (3) 이중가닥엑소뉴클레오리틱(exonucleolytic) 활성을갖는효소로상기사용프로브를분해(digesting)하여, 검출가능한상태의상기검출가능요소들및 적어도부분적으로상기제1 올리고뉴클레오티드의서열보체인단일가닥제4 올리고뉴클레오티드를생성하는단계; (4) 상기제4 올리고뉴클레오티드를다른제1 올리고뉴클레오티드와반응시켜상기사용프로브에상응하는실질적인이중가닥올리고뉴클레오티드생성물을생성하는단계; (5) 상기단계 (3) 및 (4)를주기적으로반복하여수행하는단계; 및 (6) 상기단계 (3)의매 반복마다방출되는상기특이적검출가능요소들을검출하는단계를포함하는것을특징으로한다. 바람직하게는, 검출가능요소들은형광단이다. 본발명의방법은종래개시된방법보다단일뉴클레오시드 3인산으로부터더 강한형광신호를생성한다. 또한, 바람직한프로브시스템역시개시한다.
    • 并提供了测序核酸如DNA或RNA的方法。 该实例包括(1)通过核酸的渐进焦磷酸分解产生单个核苷三磷酸酯物流; (2)提高的至少一个实质性的双链由单个三磷酸核苷中的至少一种反应并在聚合酶和连接酶的存在下相应探头系统以产生核苷酸探针使用(使用探针),所述 相应的探测系统,其中,(a)的单链标记在非检测状态的第一寡核苷酸和(b)所述第一寡核苷酸的具体可检测元件的单链,可以杂交的互补区上的核苷酸第二mitje 3个寡核苷酸; (3)的两倍链外切新克利鸭蜱(核酸外切)用具有活性的酶与所述的检测状态检测元件至第一分解(消化)所使用的探针寡核苷酸,和核苷酸序列的补体的至少一部分 产生单链第四寡核苷酸; (4)使所述第四寡核苷酸与另一个第一寡核苷酸反应以产生对应于所述使用探针的基本双链寡核苷酸产物; (5)周期性地重复上述步骤(3)和(4); 并且(6)检测步骤(3)的每次重复发出的特定可检测元素。 优选地,可检测元件是荧光团。 与先前公开的方法相比,本发明的方法从单个核苷三磷酸产生更强的荧光信号。 还公开了一种优选的探针系统。
    • 5. 发明公开
    • 단일 뉴클레오티드 검출 방법
    • 单核心检测方法
    • KR1020170018066A
    • 2017-02-15
    • KR1020177001507
    • 2015-07-22
    • 베이스4 이노베이션 엘티디
    • 밤포스,바너비프레일링,카메론알렉산더
    • C12Q1/68
    • C12Q1/6869C12Q1/6823C12Q2521/101C12Q2521/319C12Q2521/501C12Q2525/125C12Q2525/307C12Q2533/107C12Q2537/149C12Q2537/162C12Q2563/107C12Q2563/159C12Q2565/629
    • DNA 또는 RNA와같은핵산의염기서열분석방법을제공한다. 본실시예는 (1) 상기핵산의점진적인피로인산분해에의해단일뉴클레오시드 3인산스트림을생성하는단계; (2) 중합효소및 리가아제의존재하에적어도하나의상기단일뉴클레오시드 3인산과상응하는프로브시스템을반응시킴으로써적어도하나의실질적인이중가닥올리고뉴클레오티드사용프로브(used probe)를생성하는단계로, 상기상응하는프로브시스템은, (a) 검출불가능한상태의특이적검출가능요소들로표지된단일가닥제1 올리고뉴클레오티드및 (b) 상기제1 올리고뉴클레오티드상의상보적영역에혼성화할수 있는단일가닥제2 및제3 올리고뉴클레오티드를포함하는단계; (3) 이중가닥엑소뉴클레오리틱(exonucleolytic) 활성을갖는효소로상기사용프로브를분해(digesting)하여, 검출가능한상태의상기검출가능요소들및 적어도부분적으로상기제1 올리고뉴클레오티드의서열보체인단일가닥제4 올리고뉴클레오티드를생성하는단계; (4) 상기제4 올리고뉴클레오티드를다른제1 올리고뉴클레오티드와반응시켜상기사용프로브에상응하는실질적인이중가닥올리고뉴클레오티드생성물을생성하는단계; (5) 상기단계 (3) 및 (4)를주기적으로반복하여수행하는단계; 및 (6) 상기단계 (3)의매 반복마다방출되는상기특이적검출가능요소들을검출하는단계를포함하는것을특징으로한다. 바람직하게는, 검출가능요소들은형광단이다. 본발명의방법은종래개시된방법보다단일뉴클레오시드 3인산으로부터더 강한형광신호를생성한다. 또한, 바람직한프로브시스템역시개시한다.
    • 一种分析单个核苷三磷酸的方法,其特征在于,其包括以下步骤:(1)通过在聚合酶和连接酶的存在下反应制备至少一种基本上双链寡核苷酸的探针,将待捕获的单核苷三磷酸 相应的探针系统,其包含(a)以不可检测状态的特征可检测元件标记的第一单链寡核苷酸和(b)能够与第一寡核苷酸上的互补区杂交的第二和第三单链寡核苷酸; (2)用具有双链外切核酸酶活性的酶消化所使用的探针以产生可检测状态的可检测元件和至少部分为第一寡核苷酸序列互补序列的单链第四寡核苷酸; (3)使第四寡核苷酸与另一个第一寡核苷酸反应,以产生对应于所用探针的基本双链寡核苷酸产物; (4)在一个循环中重复步骤(2)和(3),(5)检测在步骤(4)的每个迭代中释放的特征可检测元素。 本发明的方法从单个核苷三磷酸产生比之前描述的更强的荧光信号。 还公开了合适的探针系统。
    • 9. 发明公开
    • 폴리뉴클레오티드를 이용한 제품 인증 장치 및 방법
    • 用于使用多核苷酸验证产品的装置和方法
    • KR1020110078185A
    • 2011-07-07
    • KR1020090134927
    • 2009-12-30
    • 삼성전자주식회사
    • 이주원박경희
    • C12Q1/68C12N15/11G06K7/00
    • C12Q1/6816B01L3/502707C12M3/00C12Q1/68C12Q1/6806C12Q1/6813C12Q1/6844C12Q2531/125C12Q2533/107C12Q2563/173
    • PURPOSE: An apparatus and method for product verification using a polynucleotide is provided to easily distinguish genuine articles and counterfeit articles. CONSTITUTION: A method for product verification comprises: a step of hybridizing a probe polynucleotide and a target polynucleotide; a step of linking the ends of a first segment and a second segment on the probe polynucleotide; a step of amplifying the probe polynucleotide; and a step of detecting the amplified product. The amplification is performed by rolling circle amplification or multiple displacement amplification. An apparatus for product verification comprises: a sample injection part(10) containing the prove polynucleotide and ligase; an amplification part(20) containing polynucleotide polymerase, primer, and dNTP; and a detection part for detecting the amplified polynucleotide.
    • 目的:提供一种使用多核苷酸进行产品验证的设备和方法,可轻松区分真品和假冒物品。 构成:产品验证的方法包括:将探针多核苷酸和靶多核苷酸杂交的步骤; 将探针多核苷酸上的第一片段和第二片段的末端连接的步骤; 扩增探针多核苷酸的步骤; 以及检测扩增产物的步骤。 通过滚环放大或多位置扩增进行扩增。 一种用于产品验证的装置,包括:含有证明多核苷酸和连接酶的样品注射部分(10) 包含多聚核苷酸聚合酶,引物和dNTP的扩增部分(20); 和用于检测扩增的多核苷酸的检测部分。
    • 10. 发明公开
    • 비드-기초 서열화를 위한 시약, 방법, 및 라이브러리
    • 用于基于BEAD的测序的试剂,方法和图谱
    • KR1020070112785A
    • 2007-11-27
    • KR1020077019819
    • 2006-02-01
    • 에이젠코트 바이오사이언스 코오포레이션
    • 맥커난,케빈블란차드,알란코틀러,레브코스타,지나
    • C12Q1/68G06F19/28
    • C12Q1/6874B82Y15/00B82Y30/00C12Q1/68C12Q1/6837C12Q1/6844C12Q1/6869C12Q2533/107C12Q2537/165C12Q2565/102C12Q2565/137C12Q2565/513C12Q2565/518C12Q2565/537G06F19/28
    • The present invention provides methods for determining a nucleic acid sequence by performing successive cycles of duplex extension along a single stranded template. The cycles comprise steps of extension, ligation, and, preferably, cleavage. In certain embodiments the methods make use of extension probes containing phosphorothiolate linkages and employ agents appropriate to cleave such linkages. In certain embodiments the methods make use of extension probes containing an abasic residue or a damaged base and employ agents appropriate to cleave linkages between a nucleoside and an abasic residue and/or agents appropriate to remove a damaged base from a nucleic acid. The invention provides methods of determining information about a sequence using at least two distinguishably labeled probe families. In certain embodiments the methods acquire less than 2 bits of information from each of a plurality of nucleotides in the template in each cycle. In certain embodiments the sequencing reactions are performed on templates attached to beads, which are immobilized in or on a semi-solid support. The invention further provides sets of labeled extension probes containing phosphorothiolate linkages or trigger residues that are suitable for use in the method. In addition, the invention includes performing multiple sequencing reactions on a single template by removing initializing oligonucleotides and extended strands and performing subsequent reactions using different initializing oligonucleotides. The invention further provides efficient methods for preparing templates, particularly for performing sequencing multiple different templates in parallel. The invention also provides methods for performing ligation and cleavage. The invention also provides new libraries of nucleic acid fragments containing paired tags, and methods of preparing microparticles having multiple different templates (e.g., containing paired tags) attached thereto and of sequencing the templates individually. The invention also provides automated sequencing systems, flow cells, image processing methods, and computer-readable media that store computer-executable instructions (e.g., to perform the image-processing methods) and/or sequence information. In certain embodiments the sequence information is stored in a database.
    • 本发明提供了通过沿着单链模板进行连续循环的双链延伸来确定核酸序列的方法。 循环包括延伸,连接和优选切割的步骤。 在某些实施方案中,所述方法利用含有硫代磷酸酯键的延伸探针,并使用适合切割这种连接的试剂。 在某些实施方案中,所述方法利用含有无碱基残基或受损碱基的延伸探针,并使用适于切割核苷和脱碱基残基之间的连接的试剂和/或适于从核酸中除去受损碱基的试剂。 本发明提供使用至少两个可区分标记的探针家族确定关于序列的信息的方法。 在某些实施方案中,该方法在每个周期中从模板中的多个核苷酸中的每一个获取少于2位的信息。 在某些实施方案中,测序反应在附着于珠上的模板上进行,所述模板固定在半固体支持物中或半固体支持物上。 本发明进一步提供了包含适合用于该方法的硫代磷酸酯键或触发性残基的标记的延伸探针的集合。 此外,本发明包括通过除去初始化寡核苷酸和延伸的链并使用不同的初始化寡核苷酸进行后续反应,在单个模板上进行多个测序反应。 本发明还提供了用于制备模板的有效方法,特别是用于并行地进行多个不同模板的排序。 本发明还提供了进行连接和切割的方法。 本发明还提供了含有配对标签的核酸片段的新文库,以及制备具有多个不同模板(例如,含有配对标签)的微粒和分别对模板进行测序的方法。 本发明还提供自动排序系统,流动池,图像处理方法和存储计算机可执行指令(例如,执行图像处理方法)和/或序列信息的计算机可读介质。 在某些实施例中,序列信息存储在数据库中。