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1. 发明申请

WO0123612A2 METHODS AND KITS FOR DETERMINING MUTATIONS, LOCAL CONFORMATIONAL CHANGES, AND AMOUNTS OF NUCLEIC ACIDS BY SELECTIVE REACTION OF THE 2'-RIBOSE POSITION IN HYBRIDIZED OLIGONUCLEOTIDES 审中-公开
标题翻译：用于通过选择性反应混合寡核苷酸中2'-位置位置来确定突变，局部一致性变化和核酸含量的方法和试剂盒
公开(公告)号：WO0123612A2
公开(公告)日：2001-04-05
申请号：PCT/US0026320
申请日：2000-09-25
申请人： UNIV NORTH CAROLINA , WEEKS KEVIN , CHAMBERLIN STACY I , JOHN DEBORAH M
发明人： WEEKS KEVIN , CHAMBERLIN STACY I , JOHN DEBORAH M
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6827
摘要： New chemical tagging methods and kits of the present invention utilize the discovery that chemical modification ( e.g. , acylation) of 2'-substituted ribonucleotides or deoxyribonucleotides by reactive compounds ( e.g. , activated esters) is sensitive to the base-paired state of the nucleotide. Perfectly base-paired positions are generally unreactive, while mismatched or unmatched bases are reactive under a wide variety of reaction conditions. The methods of the invention include a method of detecting a mutation in a nucleic acid molecule suspected of containing a mutation. The nucleic acid is hybridized to an oligonucleotide having a sequence complementary to the sequence the nucleic acid would have if a mutation were not present. The oligonucleotide comprises at least one nucleotide with a substitution at the 2'-ribose position. After the oligonucleotide is hybridized to the nucleic acid molecule, the hybridized oligonucleotide is then contacted with a reactive compound comprising a reporter moiety. Detection of the binding of the reporter moiety to the hybridized oligonucleotide indicates a mismatch i.e. , a mutation) in the nucleic acid molecule. The methods and kits of the present invention are useful in detecting point mutations and other defects in nucleic acid sequences. These methods and kits are also useful for detecting single nucleotide polymorphisms (SNPs) and mutations responsible for cancer and other genetic diseases in humans; for quantifying amounts of nucleic acids; and for detecting conformational changes in nucleic acid structures, including those found in the products of in vitro selection experiments i.e. , aptamers).
摘要翻译：本发明的新的化学标记方法和试剂盒利用这样的发现，即通过反应性化合物（例如，活化的）化学修饰（例如，酰化）2'-取代的核糖核苷酸或脱氧核糖核苷酸酯）对核苷酸的碱基配对状态敏感。完全碱基配对的位置通常是不反应的，而不匹配或不匹配的碱基在多种反应条件下是反应性的。本发明的方法包括检测疑似含有突变的核酸分子中的突变的方法。如果不存在突变，则核酸与具有与核酸将具有的序列互补的序列的寡核苷酸杂交。寡核苷酸包含至少一个在2'-核糖位置具有取代的核苷酸。寡核苷酸与核酸分子杂交后，然后将杂交的寡核苷酸与包含报道部分的反应性化合物接触。报道部分与杂交寡核苷酸结合的检测表明核酸分子中的错配（即突变）。本发明的方法和试剂盒可用于检测核酸序列中的点突变和其他缺陷。这些方法和试剂盒也可用于检测负责人类癌症和其他遗传疾病的单核苷酸多态性（SNPs）和突变; 用于量化核酸量; 并且用于检测核酸结构中的构象变化，包括在体外选择实验，适配子的产物中发现的构象变化。

2. 发明申请

WO0123612A9 DETERMINING MUTATIONS BY SELECTIVE REACTION OF THE 2'-RIBOSE POSITION IN HYBRIDIZED OLIGONUCLEOTIDES 审中-公开
标题翻译：通过选择性反应混合寡核苷酸中的2'-位置决定突变
公开(公告)号：WO0123612A9
公开(公告)日：2002-10-03
申请号：PCT/US0026320
申请日：2000-09-25
申请人： UNIV NORTH CAROLINA , WEEKS KEVIN , CHAMBERLIN STACY I , JOHN DEBORAH M
发明人： WEEKS KEVIN , CHAMBERLIN STACY I , JOHN DEBORAH M
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6827
摘要： New chemical tagging methods and kits of the present invention utilize the discovery that chemical modification ( e.g. , acylation) of 2'-substituted ribonucleotides or deoxyribonucleotides by reactive compounds ( e.g. , activated esters) is sensitive to the base-paired state of the nucleotide. Perfectly base-paired positions are generally unreactive, while mismatched or unmatched bases are reactive under a wide variety of reaction conditions. The methods of the invention include a method of detecting a mutation in a nucleic acid molecule suspected of containing a mutation. The nucleic acid is hybridized to an oligonucleotide having a sequence complementary to the sequence the nucleic acid would have if a mutation were not present. The oligonucleotide comprises at least one nucleotide with a substitution at the 2'-ribose position. After the oligonucleotide is hybridized to the nucleic acid molecule, the hybridized oligonucleotide is then contacted with a reactive compound comprising a reporter moiety. Detection of the binding of the reporter moiety to the hybridized oligonucleotide indicates a mismatch i.e. , a mutation) in the nucleic acid molecule. The methods and kits of the present invention are useful in detecting point mutations and other defects in nucleic acid sequences. These methods and kits are also useful for detecting single nucleotide polymorphisms (SNPs) and mutations responsible for cancer and other genetic diseases in humans; for quantifying amounts of nucleic acids; and for detecting conformational changes in nucleic acid structures, including those found in the products of in vitro selection experiments i.e. , aptamers).
摘要翻译：本发明的新化学标记方法和试剂盒利用了通过反应性化合物（例如活化酯）对2'-取代的核糖核苷酸或脱氧核糖核苷酸的化学修饰（例如酰化）对核苷酸的碱基配对状态敏感的发现。完全碱基配对的位置通常是不反应的，而不匹配或不匹配的碱基在多种反应条件下是反应性的。本发明的方法包括检测疑似含有突变的核酸分子中的突变的方法。如果不存在突变，则核酸与具有与核酸将具有的序列互补的序列的寡核苷酸杂交。寡核苷酸包含至少一个在2'-核糖位置具有取代的核苷酸。寡核苷酸与核酸分子杂交后，然后将杂交的寡核苷酸与包含报道部分的反应性化合物接触。检测报道部分与杂交寡核苷酸的结合表示核酸分子中的错配即突变）。本发明的方法和试剂盒可用于检测核酸序列中的点突变和其他缺陷。这些方法和试剂盒也可用于检测负责人类癌症和其他遗传疾病的单核苷酸多态性（SNPs）和突变; 用于量化核酸量; 并用于检测核酸结构中的构象变化，包括在体外选择实验的产物（即适体）中发现的结构变化。

3. 发明申请

WO2001023612A3 DETERMINING MUTATIONS BY SELECTIVE REACTION OF THE 2'-RIBOSE POSITION IN HYBRIDIZED OLIGONUCLEOTIDES 审中-公开
公开(公告)号：WO2001023612A3
公开(公告)日：2001-04-05
申请号：PCT/US2000/026320
申请日：2000-09-25
申请人： THE UNIVERSITY OF NORTH CAROLINA AT CHAPEL HILL , WEEKS, Kevin , CHAMBERLIN, Stacy, I. , JOHN, Deborah, M.
发明人： WEEKS, Kevin , CHAMBERLIN, Stacy, I. , JOHN, Deborah, M.
IPC分类号： C12Q1/68
摘要： New chemical tagging methods and kits of the present invention utilize the discovery that chemical modification ( e.g. , acylation) of 2'-substituted ribonucleotides or deoxyribonucleotides by reactive compounds ( e.g. , activated esters) is sensitive to the base-paired state of the nucleotide. Perfectly base-paired positions are generally unreactive, while mismatched or unmatched bases are reactive under a wide variety of reaction conditions. The methods of the invention include a method of detecting a mutation in a nucleic acid molecule suspected of containing a mutation. The nucleic acid is hybridized to an oligonucleotide having a sequence complementary to the sequence the nucleic acid would have if a mutation were not present. The oligonucleotide comprises at least one nucleotide with a substitution at the 2'-ribose position. After the oligonucleotide is hybridized to the nucleic acid molecule, the hybridized oligonucleotide is then contacted with a reactive compound comprising a reporter moiety. Detection of the binding of the reporter moiety to the hybridized oligonucleotide indicates a mismatch i.e. , a mutation) in the nucleic acid molecule. The methods and kits of the present invention are useful in detecting point mutations and other defects in nucleic acid sequences. These methods and kits are also useful for detecting single nucleotide polymorphisms (SNPs) and mutations responsible for cancer and other genetic diseases in humans; for quantifying amounts of nucleic acids; and for detecting conformational changes in nucleic acid structures, including those found in the products of in vitro selection experiments i.e. , aptamers).

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