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1. 发明申请

WO2004067726A2 ISOTHERMAL REACTIONS FOR THE AMPLIFICATION OF OLIGONUCLEOTIDES 审中-公开
标题翻译：用于扩增寡核苷酸的同位素反应
公开(公告)号：WO2004067726A2
公开(公告)日：2004-08-12
申请号：PCT/US2004/002718
申请日：2004-01-29
申请人： KECK GRADUATE INSTITUTE , IONIAN TECHNOLOGIES INC. , VAN NESS, Jeffrey , GALAS, David, J. , VAN NESS, Lori, K. , GRAYBILL, Jeffrey, B.
发明人： VAN NESS, Jeffrey , GALAS, David, J. , VAN NESS, Lori, K. , GRAYBILL, Jeffrey, B.
IPC分类号： C12N
CPC分类号： C12Q1/6848 , C12Q2527/101 , C12Q2525/131 , C12Q2521/301
摘要： Described is a new class of isothermal reactions for amplifying DNA. These homogeneous reactions rapidly synthesize short oligonucleotides (e.g., 8-16 bases) specified by the sequence of an amplification template. Versions of the reactions can proceed in either a linear or an exponential amplification mode. Both of these reactions utilize simple, stable conditions. The rate of amplification depends entirely on the molecular parameters governing the interactions of the molecules in the reaction. The exponential version of the method is a molecular chain reaction that uses the oligonucleotide products of each linear reaction to create a producer of more of the same oligonucleotide. It is a highly sensitive chain reaction that can be triggered by specific DNA sequences and can achieve amplifications of more than 106 fold. Several similar reactions in this class are described here. The robustness, speed and sensitivity of the exponential reaction render it useful in rapidly detecting the presence of small amounts of a specific DNA sequence in a sample, and a range of other applications including many currently making use of the polymerase chain reaction (PCR).
摘要翻译：描述了一类用于扩增DNA的等温反应。这些均相反应快速合成由扩增模板的序列指定的短寡核苷酸（例如8-16个碱基）。反应的版本可以以线性或指数放大模式进行。这两种反应都使用简单，稳定的条件。扩增速率完全取决于控制反应中分子相互作用的分子参数。该方法的指数版本是分子链反应，其使用每个线性反应的寡核苷酸产物来产生更多相同寡核苷酸的生产者。它是一种高度敏感的连锁反应，可以由特定的DNA序列引发，可以达到超过106倍的扩增。这里描述了这个类中的几个类似的反应。指数反应的鲁棒性，速度和灵敏度使其可用于快速检测样品中少量特定DNA序列的存在，以及包括目前使用聚合酶链反应（PCR）的许多其他应用的范围。

2. 发明申请

WO2004067764A2 NUCLEIC ACID SEQUENCING USING NICKING AGENTS 审中-公开
标题翻译：用酸化剂代替核酸序列
公开(公告)号：WO2004067764A2
公开(公告)日：2004-08-12
申请号：PCT/US2004/002719
申请日：2004-01-29
申请人： KECK GRADUATE INSTITUTE , VAN NESS, Jeffrey , GALAS, David, J. , VAN NESS, Lori, K.
发明人： VAN NESS, Jeffrey , GALAS, David, J. , VAN NESS, Lori, K.
IPC分类号： C12Q
CPC分类号： C12Q1/6869 , C12Q1/6872 , C12Q2533/101 , C12Q2521/313 , C12Q2521/301 , C12Q2531/101 , C12Q2525/131
摘要： Nucleotide base sequence information is obtained upon combining a template nucleic acid comprising a primer strand and a target nucleic acid strand, the target nucleic acid strand comprising an interrogation sequence, with a nicking agent that recognizes a double-stranded sequence present in the template nucleic acid and then nicks the primer strand of the template nucleic acid at a location relative to the nicking agent recognition sequence, with a distributive polymerase that will extend the primer strand from the site where the nicking agent nicked the primer strand, in the presence of suitably-reactive mononucleotides, to provide a plurality of oligonucleotides that are complements of the sequence present in the interrogation sequence, where the plurality differ in having different nucleotide lengths, so the aligning the plurality of oligonucleotides provides nucleotide base sequence information about the interrogation sequence.
摘要翻译：将包含引物链和靶核酸链的模板核酸（包括询问序列的靶核酸链）与识别模板核酸中存在的双链序列的切口剂组合时获得核苷酸碱基序列信息然后在相对于切口剂识别序列的位置处切割模板核酸的引物链，使用分布式聚合酶，其在适当的条件下存在下将延伸引物链从切口切口引物链的位点延伸，反应性单核苷酸，以提供作为询问序列中存在的序列的互补序列的多个寡核苷酸，其中多个核苷酸长度不同，因此对齐多个寡核苷酸提供了关于询问序列的核苷酸碱基序列信息。

3. 发明申请

WO2003008624A2 NUCLEIC ACID AMPLIFICATION USING NICKING AGENTS 审中-公开
标题翻译：使用镍原子的核酸放大
公开(公告)号：WO2003008624A2
公开(公告)日：2003-01-30
申请号：PCT/US2002/022748
申请日：2002-07-15
申请人： KECK GRADUATE INSTITUTE , VAN NESS, Jeffrey , GALAS, David, J. , VAN NESS, Lori, K.
发明人： VAN NESS, Jeffrey , GALAS, David, J. , VAN NESS, Lori, K.
IPC分类号： C12Q
CPC分类号： C12Q1/6844 , B01J2219/00608 , B01J2219/0061 , B01J2219/00659 , B01J2219/00722 , C12Q2600/158 , C40B40/06 , C12Q2527/107 , C12Q2525/155 , C12Q2521/301 , C12Q2531/119 , C12Q2521/513 , C12Q2521/313 , C12Q2521/307 , C12Q2521/101
摘要： The present invention provides methods and kits for amplifying target nucleic acids (including whole genomes) using nicking agents. In certain aspects, the amplification does not require the use of any external oligonucleotide primers that are capable of annealing to a portion of the target nucleic acid. This invention is useful in many areas such as genetic disease diagnoses, forensic analyses and paleoarcheological studies.
摘要翻译：本发明提供使用切口剂扩增靶核酸（包括全基因组）的方法和试剂盒。在某些方面，扩增不需要使用能够退火到靶核酸的一部分的任何外部寡核苷酸引物。本发明在诸如遗传疾病诊断，法医分析和古生物学研究等许多领域中是有用的。

4. 发明申请

WO2004067765A2 ORGANISM FINGERPRINTING USING NICKING AGENTS 审中-公开
标题翻译：有机体用指甲剂指示
公开(公告)号：WO2004067765A2
公开(公告)日：2004-08-12
申请号：PCT/US2004/002720
申请日：2004-01-29
申请人： KECK GRADUATE INSTITUTE , VAN NESS, Jeffrey , GALAS, David, J. , VAN NESS, Lori, K.
发明人： VAN NESS, Jeffrey , GALAS, David, J. , VAN NESS, Lori, K.
IPC分类号： C12Q
CPC分类号： C12Q1/6888 , C12Q2600/156 , C12Q2600/16
摘要： A sample of nucleic acid that is characteristic of a particular source, e.g., a bacterium, can be analyzed to determine whether that particular sample is from a particular source. The analysis protocol subjects the sample to a nicking reaction to produce a family of nicked fragments. In certain embodiments, the nicking reaction is performed in the presence of a polymerase and nucleoside triphosphates to amplify nicked fragments or portions thereof. The nicked fragments or portions thereof may be analyzed to determine the source of the nucleic acid sample.
摘要翻译：可以分析特定来源（例如细菌）特征的核酸样品，以确定该特定样品是否来自特定来源。分析方案使样品进行切口反应以产生切口片段家族。在某些实施方案中，切口反应在聚合酶和核苷三磷酸存在下进行以扩增切口的片段或其部分。可以分析切口的片段或其部分以确定核酸样品的来源。

5. 发明申请

WO2003008624A3 NUCLEIC ACID AMPLIFICATION USING NICKING AGENTS 审中-公开
公开(公告)号：WO2003008624A3
公开(公告)日：2003-01-30
申请号：PCT/US2002/022748
申请日：2002-07-15
申请人： KECK GRADUATE INSTITUTE , VAN NESS, Jeffrey , GALAS, David, J. , VAN NESS, Lori, K.
发明人： VAN NESS, Jeffrey , GALAS, David, J. , VAN NESS, Lori, K.
IPC分类号： C12P19/34
摘要： The present invention provides methods and kits for amplifying target nucleic acids (including whole genomes) using nicking agents. In certain aspects, the amplification does not require the use of any external oligonucleotide primers that are capable of annealing to a portion of the target nucleic acid. This invention is useful in many areas such as genetic disease diagnoses, forensic analyses and paleoarcheological studies.

6. 发明申请

WO2003008623A2 METHYLATION ANALYSIS USING NICKING AGENTS 审中-公开
标题翻译：使用镍剂进行甲基化分析
公开(公告)号：WO2003008623A2
公开(公告)日：2003-01-30
申请号：PCT/US2002/022661
申请日：2002-07-15
申请人： KECK GRADUATE INSTITUTE , VAN NESS, Jeffrey , GALAS, David, J. , VAN NESS, Lori, K.
发明人： VAN NESS, Jeffrey , GALAS, David, J. , VAN NESS, Lori, K.
IPC分类号： C12Q
CPC分类号： C12Q1/6844 , B01J2219/00608 , B01J2219/00621 , B01J2219/00659 , B01J2219/00722 , C12Q1/6827 , C12Q2600/156 , C12Q2600/158 , C40B40/06 , C12Q2531/119 , C12Q2521/513 , C12Q2521/313 , C12Q2527/107 , C12Q2525/155 , C12Q2521/301 , C12Q2565/501 , C12Q2525/131 , C12Q2523/125
摘要： The present invention provides methods and compositions for nucleic acid methylation analysis using nicking agents.
摘要翻译：本发明提供了使用切刻剂进行核酸甲基化分析的方法和组合物。

7. 发明申请

WO2003008642A2 AMPLIFICATION OF NUCLEIC ACID FRAGMENTS USING NICKING AGENTS 审中-公开
标题翻译：用镍剂扩增核酸片段
公开(公告)号：WO2003008642A2
公开(公告)日：2003-01-30
申请号：PCT/GB2002/003237
申请日：2002-07-15
申请人： KECK GRADUATE INSTITUTE , GOWSHALL, Jon, V. , VAN NESS, Jeffrey , GALAS, David, J. , VAN NESS, Lori, K.
发明人： VAN NESS, Jeffrey , GALAS, David, J. , VAN NESS, Lori, K.
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6844 , B01J2219/00608 , B01J2219/00621 , B01J2219/00659 , B01J2219/00722 , C12Q1/6827 , C12Q2600/156 , C12Q2600/158 , C40B40/06 , C12Q2531/119 , C12Q2521/513 , C12Q2521/313 , C12Q2527/107 , C12Q2525/155 , C12Q2521/301 , C12Q2565/501 , C12Q2525/131 , C12Q2523/125
摘要： The present invention relates to compounds, kits and methods for detecting a genetic variation in a target nucleic acid, detecting the presence, or the absence of a particular nucleic acid in a biological sample, preparing single-stranded nucleic acid probes, and detecting pre-mRNA differential splicing in a target cDNA molecule or a cDNA population. It utilizes a nicking agent in the amplification of a single-stranded nucleic acid fragment that either contains a genetic variation of the target nucleic acid, has a unique sequence correlating to the particular nucleic acid, is complementary to a nucleic acid of interest, or comprises an exon-exon junction. Detection and/or characterization of this short single-stranded nucleic acid fragment identifies the genetic variation of the target nucleic acid, indicates the presence of the particular nucleic acid in the sample, makes single-stranded nucleic acid probes for the nucleic acid of interest, or detecting the presence of the exon-exon junction in the target cDNA molecule or the cDNA probes.
摘要翻译：本发明涉及用于检测靶核酸中的遗传变异，检测生物样品中特定核酸的存在或不存在，制备单链的化合物，试剂盒和方法核酸探针，并检测靶cDNA分子或cDNA群体中的前体mRNA差异剪接。它在扩增单链核酸片段中使用切割剂，所述单链核酸片段包含靶核酸的遗传变异，具有与特定核酸相关的独特序列，与感兴趣的核酸互补或包含外显子 - 外显子连接。该短单链核酸片段的检测和/或表征鉴定靶核酸的遗传变异，指示样品中特定核酸的存在，针对感兴趣的核酸制备单链核酸探针，或检测目标cDNA分子或cDNA探针中外显子 - 外显子连接的存在。

8. 发明申请

WO2004022701A2 EXPONENTIAL AMPLIFICATION OF NUCLEIC ACIDS USING NICKING AGENTS 审中-公开
标题翻译：用镍试剂诱导核酸的指数扩增
公开(公告)号：WO2004022701A2
公开(公告)日：2004-03-18
申请号：PCT/US2002/022677
申请日：2002-07-15
申请人： KECK GRADUATE INSTITUTE , VAN NESS, Jeffrey , GALAS, David, J. , VAN NESS, Lori, K.
发明人： VAN NESS, Jeffrey , GALAS, David, J. , VAN NESS, Lori, K.
IPC分类号： C12N
CPC分类号： C12Q1/6809 , B01J2219/00608 , B01J2219/0061 , B01J2219/00612 , B01J2219/00621 , B01J2219/00626 , B01J2219/0063 , B01J2219/00637 , B01J2219/00659 , B01J2219/00722 , C12Q1/6844 , C12Q2600/156 , C12Q2600/158 , C40B40/06 , C12Q2525/131 , C12Q2521/313 , C12Q2565/501 , C12Q2527/107 , C12Q2525/155 , C12Q2521/301 , C12Q2531/119 , C12Q2521/513
摘要： The present invention provides methods and compositions for exponential amplification of nucleic acid molecules using nicking agents. In certain aspects, the amplification may be performed isothermally. This invention is useful in many areas such as disease diagnosis.
摘要翻译：本发明提供了使用切刻剂指数扩增核酸分子的方法和组合物。在某些方面，可以等温地进行扩增。本发明在诸如疾病诊断等许多领域是有用的。

9. 发明申请

WO2003080645A2 AMPLIFICATION OF NUCLEIC ACID FRAGMENTS USING NICKING AGENTS 审中-公开
标题翻译：用酸化剂代替核酸片段
公开(公告)号：WO2003080645A2
公开(公告)日：2003-10-02
申请号：PCT/US2002/022674
申请日：2002-07-15
申请人： KECK GRADUATE INSTITUTE , VAN NESS, Jeffrey , GALAS, David, J. , VAN NESS, Lori, K.
发明人： VAN NESS, Jeffrey , GALAS, David, J. , VAN NESS, Lori, K.
IPC分类号： C07K
CPC分类号： C12Q1/6844 , B01J2219/00608 , B01J2219/00621 , B01J2219/00659 , B01J2219/00722 , C12Q1/6827 , C12Q2600/156 , C12Q2600/158 , C40B40/06 , C12Q2531/119 , C12Q2521/513 , C12Q2521/313 , C12Q2527/107 , C12Q2525/155 , C12Q2521/301 , C12Q2565/501 , C12Q2525/131 , C12Q2523/125
摘要： The present invention relates to compounds, kits and methods for detecting a genetic variation in a target nucleic acid, detecting the presence, or the absence of a particular nucleic acid in a biological sample, preparing single-stranded nucleic acid probes, and detecting pre-mRNA differential splicing in a target cDNA molecule or a cDNA population. It utilizes a nicking agent in the amplification of a single-stranded nucleic acid fragment that either contains a genetic variation of the target nucleic acid, has a unique sequence correlating to the particular nucleic acid, is complementary to a nucleic acid of interest, or comprises an exon-exon junction. Detection and/or characterization of this short single-stranded nucleic acid fragment identifies the genetic variation of the target nucleic acid, indicates the presence of the particular nucleic acid in the sample, makes single-stranded nucleic acid probes for the nucleic acid of interest, or detecting the presence of the exon-exon junction in the target cDNA molecule or the cDNA probes.
摘要翻译：本发明涉及用于检测靶核酸的遗传变异的化合物，试剂盒和方法，检测生物样品中特定核酸的存在或不存在，制备单链核酸探针，靶cDNA分子或cDNA群体中的mRNA差异剪接。它利用切口剂在单链核酸片段的扩增中，所述单链核酸片段含有靶核酸的遗传变异，具有与特定核酸相关的唯一序列，与感兴趣的核酸互补，或包含外显子连接处。该短单链核酸片段的检测和/或表征鉴定了目标核酸的遗传变异，表明样品中特定核酸的存在，为感兴趣的核酸制备单链核酸探针，或检测靶cDNA分子或cDNA探针中外显子 - 外显子连接的存在。

10. 发明申请

WO2003066802A3 GENE EXPRESSION ANALYSIS USING NICKING AGENTS 审中-公开
公开(公告)号：WO2003066802A3
公开(公告)日：2003-08-14
申请号：PCT/US2002/022671
申请日：2002-07-15
申请人： KECK GRADUATE INSTITUTE , VAN NESS, Jeffrey , GALAS, David, J. , VAN NESS, Lori, K.
发明人： VAN NESS, Jeffrey , GALAS, David, J. , VAN NESS, Lori, K.
IPC分类号： C12Q1/68
摘要： The present invention provides methods and compositions for gene expression analyses using nicking agents.

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