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61. 发明授权

US10590453B2 Methods for amplification of nucleic acids utilizing a circularized template prepared from a target nucleic acid 有权
公开(公告)号：US10590453B2
公开(公告)日：2020-03-17
申请号：US15998679
申请日：2018-08-16
申请人： Lyle J. Arnold , Norman C. Nelson
发明人： Lyle J. Arnold , Norman C. Nelson
IPC分类号： C12P19/34 , C12Q1/6844
摘要： The present invention provides methods of amplifying a target nucleic acid utilizing a circularized template. Circularization may be achieved utilizing a bridging oligonucleotide or an inverter primer. The bridging oligonucleotide or inverted primer is extended forming a concatemeric amplicon that can then be used as a template to provide exponential amplification of the target nucleic acid.

62. 发明授权

US10174352B2 Methods for amplification of nucleic acids on solid support 有权
公开(公告)号：US10174352B2
公开(公告)日：2019-01-08
申请号：US14773362
申请日：2014-03-14
申请人： Lyle J. Arnold , Norman C. Nelson
发明人： Lyle J. Arnold , Norman C. Nelson
IPC分类号： C12Q1/68 , C12P19/34 , C12Q1/6853 , C12Q1/6865 , C07H21/02
摘要： The present invention provides methods for amplifying a nucleic acid from a sample containing a mixture of nucleic acids utilizing a solid support. Methods are provided utilizing user-defined primer oligonucleotides for directional amplification that assists in further manipulation of the target nucleic acid, such as sequencing. Methods are also provided utilizing blocker and displacer oligonucleotides for generating amplified target nucleic acids of defined length. One of these methods provides a first oligonucleotide and a second oligonucleotide affixed to a solid support or separate solid supports. The first oligonucleotide is blocked to prevent extension from the 3′-terminus and has a sequence complementary to a first portion of a target nucleic acid. The second oligonucleotide has a sequence that is identical to a second portion of the target nucleic acid. In this method, a sample is applied to the solid support and the target nucleic acid within the sample binds said first oligonucleotide. The solid support is then washed to remove unbound nucleic acids. A primer sequence containing a target binding region and a polymerase promoter sequence is then annealed to the bound target nucleic acid and extended producing a first duplex nucleic acid. The target sequence is then removed leaving a first nucleic acid that can now bind the second oligonucleotide. The second oligonucleotide is extended to produce a second duplex nucleic acid that contains a second nucleic acid. The second nucleic acid is then amplified by adding a polymerase.

63. 发明授权

US10081825B2 Methods for amplification of nucleic acids utilizing a circularized template prepared from a target nucleic acid 有权
公开(公告)号：US10081825B2
公开(公告)日：2018-09-25
申请号：US14773363
申请日：2014-03-14
申请人： Lyle J. Arnold , Norman C. Nelson
发明人： Lyle J. Arnold , Norman C. Nelson
IPC分类号： C12Q1/68 , C12P19/34 , C12Q1/6844
CPC分类号： C12P19/34 , C12Q1/6844 , C12Q2531/119 , C12Q2531/125 , C12Q2537/162
摘要： The present invention provides methods of amplifying a target nucleic acid utilizing a circularized template. Circularization may be achieved utilizing a bridging oligonucleotide or an inverter primer. The bridging oligonucleotide or inverted primer is extended forming a concatemeric amplicon that can then be used as a template to provide exponential amplification of the target nucleic acid.

64. 发明授权

US10072290B2 Methods for amplifying fragmented target nucleic acids utilizing an assembler sequence 有权
公开(公告)号：US10072290B2
公开(公告)日：2018-09-11
申请号：US14773366
申请日：2014-03-15
申请人： Lyle J. Arnold , Norman C. Nelson
发明人： Lyle J. Arnold , Norman C. Nelson
IPC分类号： C12Q1/68 , C12Q1/6858 , C12Q1/6806 , C12Q1/6844
CPC分类号： C12Q1/6858 , C12Q1/6806 , C12Q1/6844 , C12Q2525/161 , C12Q2525/186 , C12Q2525/204 , C12Q2531/101
摘要： The present invention provides methods of amplifying a fragmented target nucleic acid containing short target nucleic acid fragments utilizing an assembler sequence to convert these short fragments into longer sequences enabling their identification and interrogation. This is particularly important when attempting to identify small genetic variations, such as SNVs, present in highly fragmented nucleic acid samples. Amplification is accomplished by hybridizing the short target nucleic acid sequences to the assembler sequence, where these short sequences serve as primers for extension. Since the fragmented target nucleic acids that contain SNVs are utilized as primers on the assembler sequence they are preserved during amplification and can be detected.

65. 发明申请

US20160130623A1 Methods for Amplification of Nucleic Acids Utilizing Clamp Oligonucleotides 审中-公开
标题翻译：使用钳夹寡核苷酸扩增核酸的方法
公开(公告)号：US20160130623A1
公开(公告)日：2016-05-12
申请号：US14773365
申请日：2014-03-14
申请人： Lyle J. ARNOLD , Norman C. NELSON
发明人： Lyle J. Arnold
IPC分类号： C12P19/34
CPC分类号： C12P19/34 , C12Q1/6844 , C12Q2521/501 , C12Q2525/307 , C12Q2549/126
摘要： The present invention provides methods of amplifying a target nucleic acid utilizing a clamp oligonucleotide comprising a first target-binding region on the 3′-terminus and a second target-binding region on the 5′-terminus and tether region in between. The tether region may comprise a variety of user-defined sequences or elements that allow for further manipulation of the target nucleic acid. Such as, for example, capture followed by amplification, identification and/or sequencing. The target-binding regions bind to the target nucleic acid, the 3′-terminus functions as a primer to initiate extension across the target nucleic acid sequence and ligation of the gap results in formation of a circularized nucleic acid. This circular template can be used in a variety of processes, including amplification and sequencing.
摘要翻译：本发明提供了使用包含3'末端的第一靶结合区和5'末端之间的第二靶结合区和其间的系链区的钳位寡核苷酸来扩增靶核酸的方法。系链区域可以包含允许进一步操纵靶核酸的各种用户定义的序列或元件。例如，进行捕获，随后进行扩增，鉴定和/或测序。靶结合区域与靶核酸结合，3'-末端用作引物以引发靶核酸序列的延伸，并且间隙的连接导致形成环化的核酸。该圆形模板可用于各种过程，包括扩增和测序。

66. 发明申请

US20140329282A1 CLOSED NUCLEIC ACID STRUCTURES 审中-公开
标题翻译：封闭的核酸结构
公开(公告)号：US20140329282A1
公开(公告)日：2014-11-06
申请号：US14342764
申请日：2012-09-06
申请人： Norman C. Nelson , Jijumon Chelliserry , Steven T. Brentano , Dmitry Lyakhov , Matthew C. Friedenberg , Anne-Laure Shapiro
发明人： Norman C. Nelson , Jijumon Chelliserry , Steven T. Brentano , Dmitry Lyakhov , Matthew C. Friedenberg , Anne-Laure Shapiro
IPC分类号： C12P19/34
摘要： The invention provides compositions and methods for making closed nucleic acid structures in which one or both strands are continuous. The closed nucleic acid structures can be used as sequencing templates among other applications.
摘要翻译：本发明提供用于制备其中一条或两条链是连续的封闭核酸结构的组合物和方法。封闭的核酸结构可以用作其他应用中的测序模板。

67. 发明授权

US08480976B2 Instruments and methods for mixing the contents of a detection chamber 有权
标题翻译：用于混合检测室内容物的仪器和方法
公开(公告)号：US08480976B2
公开(公告)日：2013-07-09
申请号：US13181850
申请日：2011-07-13
申请人： Scott S. Breidenthal , Sara H. Fan , Richard S. Lee , Norman C. Nelson , Matthew J. Scott , Jason A. Taylor
发明人： Scott S. Breidenthal , Sara H. Fan , Richard S. Lee , Norman C. Nelson , Matthew J. Scott , Jason A. Taylor
IPC分类号： B01L3/00
CPC分类号： B01F11/00 , B01F3/1271 , B01F11/0045 , B01F11/0071 , B01F13/0059 , B01F15/00214 , B01F2215/0037 , B01L3/502 , B01L3/502715 , B01L3/50273 , B01L3/502761 , B01L7/52 , B01L7/525 , B01L2200/0621 , B01L2200/0647 , B01L2200/0673 , B01L2200/0689 , B01L2200/10 , B01L2200/14 , B01L2200/16 , B01L2300/0816 , B01L2300/087 , B01L2300/0887 , B01L2300/123 , B01L2300/1805 , B01L2300/1822 , B01L2300/1883 , B01L2400/0457 , B01L2400/0478 , B01L2400/0481 , B01L2400/0655 , B01L2400/0677 , C12N15/1006 , C12Q1/6806 , C12Q1/6844 , G01N1/44 , G01N21/01 , G01N21/6428 , G01N21/645 , G01N35/0098 , G01N2021/6419 , G01N2021/6421 , G01N2021/6432 , G01N2021/6439 , G01N2021/6463 , G01N2035/00544 , G01N2201/0627 , Y10T137/0391 , Y10T436/143333 , Y10T436/25 , Y10T436/255
摘要： A receptacle having a plurality of interconnected chambers arranged to permit multiple process steps or processes to be performed independently or simultaneously. The receptacles are manufactured to separate liquid from dried reagents and to maintain the stability of the dried reagents. An immiscible liquid, such as an oil, is included to control loading of process materials, facilitate mixing and reconstitution of dried reagents, limit evaporation, control heating of reaction materials, concentrate solid support materials to prevent clogging of fluid connections, provide minimum volumes for fluid transfers, and to prevent process materials from sticking to chamber surfaces. The receptacles can be adapted for use in systems having a processing instrument that includes an actuator system for selectively moving fluid substances between chambers and a detector. The actuator system can be arranged to concentrate an analyte present in a sample. The detector can be used to detect an optical signal emitted by the contents of the receptacle.
摘要翻译：具有多个互连室的容器，其布置成允许独立地或同时地执行多个工艺步骤或工艺。制造容器以将液体与干燥试剂分开并保持干燥试剂的稳定性。包括不混溶的液体，如油，以控制加工材料的加载，促进干燥试剂的混合和重构，限制蒸发，控制反应材料的加热，浓缩固体支持材料以防止流体连接堵塞，为流体转移，并防止工艺材料粘附到室表面。插座可以适用于具有处理器具的系统，该处理器具包括用于选择性地移动腔室和检测器之间的流体物质的致动器系统。致动器系统可以被布置成集中存在于样品中的分析物。检测器可用于检测由容器的内容物发射的光信号。

68. 发明申请

US20120264122A1 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：US20120264122A1
公开(公告)日：2012-10-18
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.
发明人： Steven T. BRENTANO , Dmitry LYAKHOV , James D. CARLSON , Norman C. NELSON , Lyle J. ARNOLD, JR.
IPC分类号： C12P19/34 , C12Q1/68 , C07H21/04
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。

69. 发明申请

US20120231456A1 INSTRUMENTS AND METHODS FOR MIXING THE CONTENTS OF A DETECTION CHAMBER 有权
标题翻译：用于混合检测室内容的仪器和方法
公开(公告)号：US20120231456A1
公开(公告)日：2012-09-13
申请号：US13414195
申请日：2012-03-07
申请人： Scott S. Breidenthal , Sara H. Fan , Richard S. Lee , Norman C. Nelson , Matthew J. Scott , Jason A. Taylor
发明人： Scott S. Breidenthal , Sara H. Fan , Richard S. Lee , Norman C. Nelson , Matthew J. Scott , Jason A. Taylor
IPC分类号： G01N21/64
CPC分类号： B01F11/00 , B01F3/1271 , B01F11/0045 , B01F11/0071 , B01F13/0059 , B01F15/00214 , B01F2215/0037 , B01L3/502 , B01L3/502715 , B01L3/50273 , B01L3/502761 , B01L7/52 , B01L7/525 , B01L2200/0621 , B01L2200/0647 , B01L2200/0673 , B01L2200/0689 , B01L2200/10 , B01L2200/14 , B01L2200/16 , B01L2300/0816 , B01L2300/087 , B01L2300/0887 , B01L2300/123 , B01L2300/1805 , B01L2300/1822 , B01L2300/1883 , B01L2400/0457 , B01L2400/0478 , B01L2400/0481 , B01L2400/0655 , B01L2400/0677 , C12N15/1006 , C12Q1/6806 , C12Q1/6844 , G01N1/44 , G01N21/01 , G01N21/6428 , G01N21/645 , G01N35/0098 , G01N2021/6419 , G01N2021/6421 , G01N2021/6432 , G01N2021/6439 , G01N2021/6463 , G01N2035/00544 , G01N2201/0627 , Y10T137/0391 , Y10T436/143333 , Y10T436/25 , Y10T436/255
摘要： A receptacle having interconnected chambers arranged to permit multiple process steps to be performed independently or simultaneously. The receptacles are manufactured to separate liquid from dried reagents and to maintain the stability of the dried reagents. An immiscible liquid, such as an oil, is included to control loading of process materials, facilitate mixing and reconstitution of dried reagents, limit evaporation, control heating of reaction materials, concentrate solid support materials to prevent clogging of fluid connections, provide minimum volumes for fluid transfers, and to prevent process materials from sticking to chamber surfaces. The receptacles can be adapted for use in systems having a processing instrument that includes an actuator system for selectively moving fluid substances between chambers and a detector. The actuator system can be arranged to concentrate an analyte present in a sample. The detector can be used to detect an optical signal emitted by the contents of the receptacle.
摘要翻译：具有互连室的容器，其布置成允许独立地或同时地执行多个工艺步骤。制造容器以将液体与干燥试剂分开并保持干燥试剂的稳定性。包括不混溶的液体，如油，以控制加工材料的加载，促进干燥试剂的混合和重构，限制蒸发，控制反应材料的加热，浓缩固体支持材料以防止流体连接堵塞，为流体转移，并防止工艺材料粘附到室表面。插座可以适用于具有处理器具的系统，该处理器具包括用于选择性地移动腔室和检测器之间的流体物质的致动器系统。致动器系统可以被布置成集中存在于样品中的分析物。检测器可用于检测由容器的内容物发射的光信号。

70. 发明授权

US08052929B2 Gravity-assisted mixing methods 有权
标题翻译：重力辅助混合法
公开(公告)号：US08052929B2
公开(公告)日：2011-11-08
申请号：US13078348
申请日：2011-04-01
申请人： Scott S. Breidenthal , Sara H. Fan , Richard S. Lee , Norman C. Nelson , Matthew J. Scott , Jason A. Taylor
发明人： Scott S. Breidenthal , Sara H. Fan , Richard S. Lee , Norman C. Nelson , Matthew J. Scott , Jason A. Taylor
IPC分类号： G01N15/06
CPC分类号： B01F11/00 , B01F3/1271 , B01F11/0045 , B01F11/0071 , B01F13/0059 , B01F15/00214 , B01F2215/0037 , B01L3/502 , B01L3/502715 , B01L3/50273 , B01L3/502761 , B01L7/52 , B01L7/525 , B01L2200/0621 , B01L2200/0647 , B01L2200/0673 , B01L2200/0689 , B01L2200/10 , B01L2200/14 , B01L2200/16 , B01L2300/0816 , B01L2300/087 , B01L2300/0887 , B01L2300/123 , B01L2300/1805 , B01L2300/1822 , B01L2300/1883 , B01L2400/0457 , B01L2400/0478 , B01L2400/0481 , B01L2400/0655 , B01L2400/0677 , C12N15/1006 , C12Q1/6806 , C12Q1/6844 , G01N1/44 , G01N21/01 , G01N21/6428 , G01N21/645 , G01N35/0098 , G01N2021/6419 , G01N2021/6421 , G01N2021/6432 , G01N2021/6439 , G01N2021/6463 , G01N2035/00544 , G01N2201/0627 , Y10T137/0391 , Y10T436/143333 , Y10T436/25 , Y10T436/255
摘要： A receptacle having a plurality of interconnected chambers arranged to permit multiple process steps or processes to be performed independently or simultaneously. The receptacles are manufactured to separate liquid from dried reagents and to maintain the stability of the dried reagents. An immiscible liquid, such as an oil, is included to control loading of process materials, facilitate mixing and reconstitution of dried reagents, limit evaporation, control heating of reaction materials, concentrate solid support materials to prevent clogging of fluid connections, provide minimum volumes for fluid transfers, and to prevent process materials from sticking to chamber surfaces. The receptacles can be adapted for use in systems having a processing instrument that includes an actuator system for selectively moving fluid substances between chambers and a detector.
摘要翻译：具有多个互连室的容器，其布置成允许独立地或同时地执行多个工艺步骤或工艺。制造容器以将液体与干燥试剂分开并保持干燥试剂的稳定性。包括不混溶的液体，如油，以控制加工材料的加载，促进干燥试剂的混合和重构，限制蒸发，控制反应材料的加热，浓缩固体支持材料以防止流体连接堵塞，为流体转移，并防止工艺材料粘附到室表面。插座可以适用于具有处理器具的系统，该处理器具包括用于选择性地移动腔室和检测器之间的流体物质的致动器系统。

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