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1. 发明授权

US08512955B2 Methods and compositions for nucleic acid amplification 有权
公开(公告)号：US08512955B2
公开(公告)日：2013-08-20
申请号：US12828676
申请日：2010-07-01
申请人： Steven T. Brentano , Dmitry Lyakhov , Norman C. Nelson , James D. Carlson , Michael M. Becker , Lyle J. Arnold, Jr.
发明人： Steven T. Brentano , Dmitry Lyakhov , Norman C. Nelson , James D. Carlson , Michael M. Becker , Lyle J. Arnold, Jr.
IPC分类号： C12Q1/68 , C12P19/34 , C07H21/04
CPC分类号： C12Q1/6865 , C12Q1/6844 , C12Q1/6848 , C12Q1/6853 , C12Q2549/119 , C12Q2537/143 , C12Q2531/143
摘要： Compositions, reaction mixtures, and methods for performing an amplification reaction, including multiplex amplification reaction, wherein the method comprises using one or more amplification oligomer complexes comprising linked first and second amplification oligomer members. In one aspect, the amplification oligomer complex is hybridized to a target nucleic acid, the target nucleic acid with hybridized amplification oligomer complex is then captured, and other components are washed away. Target sequences of the target nucleic acids are pre-amplified to generate a first amplification product. The first amplification product is amplified in one or more secondary amplification reactions to generate second amplification products.

2. 发明申请

US20120178636A1 METHODS AND COMPOSITIONS FOR NUCLEIC ACID AMPLIFICATION 有权
标题翻译：用于核酸扩增的方法和组合物
公开(公告)号：US20120178636A1
公开(公告)日：2012-07-12
申请号：US13380018
申请日：2010-07-01
申请人： Steven T. Brentano , Dmitry Lyakhov , James D. Carlson , Michael M. Becker , Norman C. Nelson , Lyle J. Arnold, JR.
发明人： Steven T. Brentano , Dmitry Lyakhov , James D. Carlson , Michael M. Becker , Norman C. Nelson , Lyle J. Arnold, JR.
IPC分类号： C40B30/00
CPC分类号： C12Q1/6865 , C12Q1/6844 , C12Q1/6848 , C12Q1/6853 , C12Q2549/119 , C12Q2537/143 , C12Q2531/143
摘要： Compositions, reaction mixtures, and methods for performing an amplification reaction, including multiplex amplification reaction, wherein the method comprises using one or more amplification oligomer complexes comprising linked first and second amplification oligomer members. In one aspect, the amplification oligomer complex is hybridized to a target nucleic acid, the target nucleic acid with hybridized amplification oligomer complex is then captured, and other components are washed away. Target sequences of the target nucleic acids are pre-amplified to generate a first amplification product. The first amplification product is amplified in one or more secondary amplification reactions to generate second amplification products.
摘要翻译：用于进行扩增反应的组合物，反应混合物和方法，包括多重扩增反应，其中所述方法包括使用包含连接的第一和第二扩增寡聚体成员的一种或多种扩增寡聚体复合物。一方面，将扩增寡聚物复合物与靶核酸杂交，然后捕获具有杂交扩增寡聚物复合物的靶核酸，并洗涤其它组分。靶核酸的靶序列被预扩增以产生第一扩增产物。第一扩增产物在一个或多个二级扩增反应中扩增以产生第二扩增产物。

3. 发明授权

US09169512B2 Methods and compositions for nucleic acid amplification 有权
标题翻译：用于核酸扩增的方法和组合物
公开(公告)号：US09169512B2
公开(公告)日：2015-10-27
申请号：US13380018
申请日：2010-07-01
申请人： Steven T. Brentano , Dmitry Lyakhov , James D. Carlson , Michael M. Becker , Norman C. Nelson , Lyle J. Arnold, Jr.
发明人： Steven T. Brentano , Dmitry Lyakhov , James D. Carlson , Michael M. Becker , Norman C. Nelson , Lyle J. Arnold, Jr.
IPC分类号： C12P19/34 , C12Q1/68
CPC分类号： C12Q1/6865 , C12Q1/6844 , C12Q1/6848 , C12Q1/6853 , C12Q2549/119 , C12Q2537/143 , C12Q2531/143
摘要： Compositions, reaction mixtures, and methods for performing an amplification reaction, including multiplex amplification reaction, wherein the method comprises using one or more amplification oligomer complexes comprising linked first and second amplification oligomer members. In one aspect, the amplification oligomer complex is hybridized to a target nucleic acid, the target nucleic acid with hybridized amplification oligomer complex is then captured, and other components are washed away. Target sequences of the target nucleic acids are pre-amplified to generate a first amplification product. The first amplification product is amplified in one or more secondary amplification reactions to generate second amplification products.
摘要翻译：用于进行扩增反应的组合物，反应混合物和方法，包括多重扩增反应，其中所述方法包括使用包含连接的第一和第二扩增寡聚体成员的一种或多种扩增寡聚体复合物。一方面，将扩增寡聚物复合物与靶核酸杂交，然后捕获具有杂交扩增寡聚物复合物的靶核酸，并洗涤其它组分。靶核酸的靶序列被预扩增以产生第一扩增产物。第一扩增产物在一个或多个二级扩增反应中扩增以产生第二扩增产物。

4. 发明授权

US08642268B2 Methods and compositions for nucleic acid amplification 有权
标题翻译：用于核酸扩增的方法和组合物
公开(公告)号：US08642268B2
公开(公告)日：2014-02-04
申请号：US13460341
申请日：2012-04-30
申请人： Steven T. Brentano , Dmitry Lyakhov , James D. Carlson , Norman C. Nelson , Lyle J. Arnold, Jr. , Michael M. Becker
发明人： Steven T. Brentano , Dmitry Lyakhov , James D. Carlson , Norman C. Nelson , Lyle J. Arnold, Jr. , Michael M. Becker
IPC分类号： C12Q1/68 , C12P19/34 , C07H19/00
CPC分类号： C12Q1/6881 , C12Q1/6834 , C12Q1/6844 , C12Q1/6865 , C12Q2525/155 , C12Q2525/186 , C12Q2525/197 , C12Q2531/143 , C12Q2565/543
摘要： Compositions that are used in nucleic acid amplification in vitro are disclosed, which include a target specific universal (TSU) promoter primer or promoter provider oligonucleotide that includes a target specific (TS) sequence that hybridizes specifically to a target sequence that is amplified and a universal (U) sequence that is introduced into the sequence that is amplified, by using a primer for the universal sequence. Methods of nucleic acid amplification in vitro are disclosed that use one or more TSU oligonucleotides to attached a U sequence to a target nucleic acid in a target capture step and then use a primer for a U sequence in subsequent amplification steps performed in substantially isothermal conditions to make amplification products that contain a U sequence that indicates the presence of the target nucleic acid in a sample.
摘要翻译：公开了用于体外核酸扩增的组合物，其包括目标特异性通用（TSU）启动子引物或启动子提供者寡核苷酸，其包括与扩增的靶序列特异性杂交的靶特异性（TS）序列和通用（U）序列，通过使用通用序列的引物引入扩增的序列。公开了体外核酸扩增方法，其中使用一种或多种TSU寡核苷酸将U序列连接到目标捕获步骤中的靶核酸，然后在基本上等温条件下进行的随后扩增步骤中使用U序列引物制备含有指示样品中靶核酸存在的U序列的扩增产物。

5. 发明授权

US07696337B2 Composition kits and methods for performing amplification reactions 有权
标题翻译：用于进行扩增反应的组合物试剂盒和方法
公开(公告)号：US07696337B2
公开(公告)日：2010-04-13
申请号：US11574307
申请日：2005-08-26
申请人： Michael M. Becker , Steven T. Brentano , Daniel P. Kolk , Wai-Chung Lam , Kristin W. Livezey , Norman C. Nelson , Astrid R. W. Schroder , Gary P. Schroth
发明人： Michael M. Becker , Steven T. Brentano , Daniel P. Kolk , Wai-Chung Lam , Kristin W. Livezey , Norman C. Nelson , Astrid R. W. Schroder , Gary P. Schroth
IPC分类号： C07H21/04 , C12Q1/68
CPC分类号： C12P19/34 , C12Q1/6865 , C12Q2537/163 , C12Q2521/325 , C12Q2521/107 , C12Q2525/186 , C12Q2533/101 , C12Q2525/143
摘要： The present invention is directed to novel methods of synthesizing multiple copies of a target nucleic acid sequence which are autocatalytic (i.e., able to cycle automatically without the need to modify reaction conditions such as temperature, pH, or ionic strength and using the product of one cycle in the next one). In particular, the present invention discloses a method of nucleic acid amplification which is robust and efficient, while reducing the appearance of side products. The method uses only one primer, the “priming oligonucleotide,” a 3′blocked promoter oligonucleotide and optionally, a means for terminating a primer extension reaction, to amplify RNA or DNA molecules in vitro, while reducing or eliminating the formation of side products. The method of the present invention minimizes or eliminates the emergence of side products, thus providing a high level of specificity. Furthermore, the appearance of side products can complicate the analysis of the amplification reaction by various molecular detection techniques. The present invention minimizes or eliminates this problem, thus providing an enhanced level of sensitivity.
摘要翻译：本发明涉及合成目标核酸序列的多个拷贝的新型方法，所述目标核酸序列是自动催化的（即，能够自动循环而不需要修饰诸如温度，pH或离子强度的反应条件，并且使用一种循环在下一个）。特别地，本发明公开了一种鲁棒有效的核酸扩增方法，同时减少了副产物的外观。该方法仅使用一种引物，即引物寡核苷酸，3'封闭的启动子寡核苷酸和任选的终止引物延伸反应的手段，以体外扩增RNA或DNA分子，同时减少或消除副产物的形成。本发明的方法使副产物的出现最小化或消除，从而提供高水平的特异性。此外，副产物的出现可能通过各种分子检测技术使扩增反应的分析复杂化。本发明使这个问题最小化或消除了这一点，从而提供了增强的灵敏度。

6. 发明授权

US07939260B2 Method for making available a priming oligonucleotide 有权
标题翻译：提供引物寡核苷酸的方法
公开(公告)号：US07939260B2
公开(公告)日：2011-05-10
申请号：US12389993
申请日：2009-02-20
申请人： Michael M. Becker , Steven T. Brentano , Kristin W. Livezey , Norman C. Nelson , Gary P. Schroth
发明人： Michael M. Becker , Steven T. Brentano , Kristin W. Livezey , Norman C. Nelson , Gary P. Schroth
IPC分类号： C12Q1/68 , C12P19/34
CPC分类号： C12P19/34 , C12Q1/6865 , C12Q2537/163 , C12Q2521/325 , C12Q2521/107 , C12Q2525/186 , C12Q2533/101 , C12Q2525/143
摘要： The present invention is directed to novel methods of synthesizing multiple copies of a target nucleic acid sequence which are autocatalytic (i.e., able to cycle automatically without the need to modify reaction conditions such as temperature, pH, or ionic strength and using the product of one cycle in the next one). In particular, the present invention discloses a method of nucleic acid amplification which is robust and efficient, while reducing the appearance of side-products. The method uses only one primer, the “priming oligonucleotide,” a promoter oligonucleotide modified to prevent polymerase extension from its 3′-terminus and, optionally, a means for terminating a primer extension reaction, to amplify RNA or DNA molecules in vitro, while reducing or substantially eliminating the formation of side-products. The method of the present invention minimizes or substantially eliminates the emergence of side-products, thus providing a high level of specificity. Furthermore, the appearance of side-products can complicate the analysis of the amplification reaction by various molecular detection techniques. The present invention minimizes or substantially eliminates this problem, thus providing an enhanced level of sensitivity.
摘要翻译：本发明涉及合成目标核酸序列的多个拷贝的新型方法，所述目标核酸序列是自动催化的（即，能够自动循环而不需要修饰诸如温度，pH或离子强度的反应条件，并且使用一种循环在下一个）。特别地，本发明公开了一种鲁棒有效的核酸扩增方法，同时减少副产物的外观。该方法仅使用一种引物，即引物寡核苷酸，经修饰的启动子寡核苷酸，用于防止聚合酶从其3'末端延伸，以及任选的终止引物延伸反应的手段，以在体外扩增RNA或DNA分子，同时减少或基本上消除副产物的形成。本发明的方法使副产物的出现最小化或基本上消除，从而提供高水平的特异性。此外，副产物的出现可能通过各种分子检测技术使扩增反应的分析复杂化。本发明使这个问题最小化或基本消除，从而提供了增强的灵敏度。

7. 发明授权

US07070925B1 Method for determining the presence of an RNA analyte in a sample using a modified oligonucleotide probe 有权
标题翻译：使用修饰的寡核苷酸探针测定样品中RNA分析物的存在的方法
公开(公告)号：US07070925B1
公开(公告)日：2006-07-04
申请号：US09565427
申请日：2000-05-05
申请人： Michael M. Becker , Mehrdad Majlessi , Steven T. Brentano
发明人： Michael M. Becker , Mehrdad Majlessi , Steven T. Brentano
IPC分类号： C07H22/04 , C12Q1/68
CPC分类号： C12Q1/6832 , C07H21/00 , C12Q1/6816 , C12Q1/6844 , C12Q1/686 , C12Q1/6865 , C12Q2563/131 , C12Q2563/113 , C12Q2525/101 , C12Q2525/117 , C12Q2527/107
摘要： The present invention concerns oligonucleotides containing one or more modified nucleotides which increase the binding affinity of the oligonucleotides to target nucleic acids having a complementary nucleotide base sequence. These modified oligonucleotides hybridize to the target sequence at a faster rate than unmodified oligonucleotides having an identical nucleotide base sequence. Such modified oligonucleotides include oligonucleotides containing at least one 2′-O-methylribofuranosyl moiety joined to a nitrogenous base. Oligonucleotides can be modified in accordance with the present invention to preferentially bind RNA targets. The present invention also concerns methods of using these modified oligonucleotides and kits containing the same.
摘要翻译：本发明涉及含有一个或多个修饰的核苷酸的寡核苷酸，其增加寡核苷酸与具有互补核苷酸碱基序列的靶核酸的结合亲和力。这些修饰的寡核苷酸以比具有相同核苷酸碱基序列的未修饰寡核苷酸更快的速率与靶序列杂交。这种修饰的寡核苷酸包括含有至少一个连接到含氮碱基的2'-O-甲基呋喃核糖基部分的寡核苷酸。可以根据本发明修饰寡核苷酸以优先结合RNA靶。本发明还涉及使用这些修饰的寡核苷酸的方法和含有该寡核苷酸的试剂盒。

8. 发明授权

US06534273B2 Two-step hybridization and capture of a polynucleotide 有权
公开(公告)号：US06534273B2
公开(公告)日：2003-03-18
申请号：US09956412
申请日：2001-09-18
申请人： William G. Weisburg , Jay H. Shaw , Michael M. Becker , Mehrdad R. Majlessi , Steven T. Brentano , Kiyotada Nunomura
发明人： William G. Weisburg , Jay H. Shaw , Michael M. Becker , Mehrdad R. Majlessi , Steven T. Brentano , Kiyotada Nunomura
IPC分类号： C12Q168
CPC分类号： C12Q1/6834 , C12Q1/6813
摘要： A method for capturing a target polynucleotide in a sample onto a solid support with an attached immobilized probe by using a capture probe and two different hybridization conditions that control the order of hybridization, where the first hybridization condition allows hybridization of the capture probe to the target polynucleotide, and the second hybridization condition allows hybridization of the capture probe to the immobilized probe. The method further includes amplifying the captured target polynucleotide by hybridizing at least one primer oligonucleotide to the target polynucleotide and using nucleic acid amplification that initiates from the primer oligonucleotide.

9. 发明授权

US07374885B2 Single-primer nucleic acid amplification methods 有权
标题翻译：单引物核酸扩增方法
公开(公告)号：US07374885B2
公开(公告)日：2008-05-20
申请号：US11213519
申请日：2005-08-26
申请人： Michael M. Becker , Wai-Chung Lam , Kristin W. Livezey , Steven T. Brentano , Daniel P. Kolk , Astrid R. W. Schroder
发明人： Michael M. Becker , Wai-Chung Lam , Kristin W. Livezey , Steven T. Brentano , Daniel P. Kolk , Astrid R. W. Schroder
IPC分类号： C12Q1/68 , C12P19/34
CPC分类号： C12P19/34 , C12Q1/6865 , C12Q2537/163 , C12Q2521/325 , C12Q2521/107 , C12Q2525/186 , C12Q2533/101 , C12Q2525/143
摘要： The present invention is directed to novel methods of synthesizing multiple copies of a target nucleic acid sequence which are autocatalytic (i.e., able to cycle automatically without the need to modify reaction conditions such as temperature, pH, or ionic strength and using the product of one cycle in the next one). In particular, the present invention discloses a method of nucleic acid amplification which is robust and efficient, while reducing the appearance of side-products. The method uses only one primer, the “priming oligonucleotide,” a promoter oligonucleotide modified to prevent polymerase extension from its 3′-terminus and, optionally, a means for terminating a primer extension reaction, to amplify RNA or DNA molecules in vitro, while reducing or substantially eliminating the formation of side-products. The method of the present invention minimizes or substantially eliminates the emergence of side-products, thus providing a high level of specificity. Furthermore, the appearance of side-products can complicate the analysis of the amplification reaction by various molecular detection techniques. The present invention minimizes or substantially eliminates this problem, thus providing an enhanced level of sensitivity.
摘要翻译：本发明涉及合成目标核酸序列的多个拷贝的新型方法，所述目标核酸序列是自动催化的（即，能够自动循环而不需要修饰诸如温度，pH或离子强度的反应条件，并且使用一种循环在下一个）。特别地，本发明公开了一种鲁棒有效的核酸扩增方法，同时减少副产物的外观。该方法仅使用一种引物，即引物寡核苷酸，经修饰的启动子寡核苷酸，用于防止聚合酶从其3'末端延伸，以及任选的终止引物延伸反应的手段，以在体外扩增RNA或DNA分子，同时减少或基本上消除副产物的形成。本发明的方法使副产物的出现最小化或基本上消除，从而提供高水平的特异性。此外，副产物的出现可能通过各种分子检测技术使扩增反应的分析复杂化。本发明使这个问题最小化或基本消除，从而提供了增强的灵敏度。

10. 发明授权

US06903206B1 Kits for amplifying target nucleic acid sequences using modified oligonucleotides 有权
标题翻译：使用修饰的寡核苷酸扩增靶核酸序列的试剂盒
公开(公告)号：US06903206B1
公开(公告)日：2005-06-07
申请号：US09523237
申请日：2000-03-10
申请人： Michael M. Becker , Mehrdad Majlessi , Steven T. Brentano
发明人： Michael M. Becker , Mehrdad Majlessi , Steven T. Brentano
IPC分类号： C12N15/09 , C07H21/00 , C12Q1/68 , C07H21/04
CPC分类号： C07H21/00 , C12Q1/6832 , C12Q1/686 , C12Q1/6865 , C12Q2527/107 , C12Q2525/113 , C12Q2523/113 , C12Q2537/101 , C12Q2565/107
摘要： The present invention concerns oligonucleotides containing one or more modified nucleotides which increase the binding affinity of the oligonucleotides to target nucleic acids having a complementary nucleotide base sequence. These modified oligonucleotides hybridize to the target sequence at a faster rate than unmodified oligonucleotides having an identical nucleotide base sequence. Such modified oligonucleotides include oligonucleotides containing at least one 2′-O-methylribofuranosyl moiety joined to a nitrogenous base. Oligonucleotides can be modified in accordance with the present invention to preferentially bind RNA targets. The present invention also concerns methods of using these modified oligonucleotides and kits containing the same.
摘要翻译：本发明涉及含有一个或多个修饰的核苷酸的寡核苷酸，其增加寡核苷酸与具有互补核苷酸碱基序列的靶核酸的结合亲和力。这些修饰的寡核苷酸以比具有相同核苷酸碱基序列的未修饰寡核苷酸更快的速率与靶序列杂交。这种修饰的寡核苷酸包括含有至少一个连接到含氮碱基的2'-O-甲基呋喃核糖基部分的寡核苷酸。可以根据本发明修饰寡核苷酸以优先结合RNA靶。本发明还涉及使用这些修饰的寡核苷酸的方法和含有该寡核苷酸的试剂盒。

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