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1. 发明授权

US08906831B2 Single molecule loading methods and compositions 有权
标题翻译：单分子加载方法和组成
公开(公告)号：US08906831B2
公开(公告)日：2014-12-09
申请号：US12384097
申请日：2009-03-30
申请人： John Eid , Stephen Turner , Ravi Dalal , Benjamin Flusberg , Jonas Korlach , Steven Lin , Adrian Fehr , Fred Christians , Robin Emig , Jeremy Gray , Robert Sebra , Lei Sun , Stephen Dudek
发明人： John Eid , Stephen Turner , Ravi Dalal , Benjamin Flusberg , Jonas Korlach , Steven Lin , Adrian Fehr , Fred Christians , Robin Emig , Jeremy Gray , Robert Sebra , Lei Sun , Stephen Dudek
IPC分类号： C12Q1/48 , C12N11/14 , G01N33/543 , C40B20/02 , C40B60/08 , C40B40/06 , C12Q1/68
CPC分类号： B01J19/0046 , B01J2219/00596 , C12N2310/50 , C12N2310/51 , C12Q1/6869 , C12Q2535/122 , C40B40/06 , G01N33/54313 , G01N33/54366
摘要： Methods for non-random loading of single analyte molecules into array structures are provided. The methods allow for distribution of a population of target molecules into a plurality of size confined regions such as wells. Sizing moieties are linked to individual target molecules. The sizing moieties are of sufficient size, relative to the size-confined reaction or observation regions, such that only a selected number of sizing moieties will fit into the size confined regions. The confined regions and the sizing moieties or target molecules comprise a selected charge that allow for controlling the loading of the sizing moities.
摘要翻译：提供了将单一分析物分子非随机加载到阵列结构中的方法。所述方法允许将目标分子群体分配到多个尺寸的限制区域例如孔中。上限部分与单独的靶分子连接。尺寸部分相对于尺寸限制的反应或观察区域具有足够的尺寸，使得只有选定数量的施胶部分将适合尺寸限制区域。限制区域和施胶部分或靶分子包括允许控制施胶剂的载荷的选定的电荷。

2. 发明申请

US20100009872A1 Single molecule loading methods and compositions 有权
标题翻译：单分子加载方法和组成
公开(公告)号：US20100009872A1
公开(公告)日：2010-01-14
申请号：US12384097
申请日：2009-03-30
申请人： John Eid , Stephen Turner , Ravi Dalal , Benjamin Flusberg , Jonas Korlach , Steven Lin , Adrian Fehr , Fred Christians , Robin Emig
发明人： John Eid , Stephen Turner , Ravi Dalal , Benjamin Flusberg , Jonas Korlach , Steven Lin , Adrian Fehr , Fred Christians , Robin Emig
IPC分类号： C40B50/06 , C40B50/00 , C40B50/18 , C40B60/12
CPC分类号： B01J19/0046 , B01J2219/00596 , C12N2310/50 , C12N2310/51 , C12Q1/6869 , C12Q2535/122 , C40B40/06 , G01N33/54313 , G01N33/54366
摘要： Methods, compositions and arrays for non-random loading of single analyte molecules into array structures are provided.
摘要翻译：提供了将单一分析物分子非随机加载到阵列结构中的方法，组合物和阵列。

3. 发明申请

US20090286245A1 TWO SLOW-STEP POLYMERASE ENZYME SYSTEMS AND METHODS 有权
标题翻译：两个慢步聚合酶酶系统和方法
公开(公告)号：US20090286245A1
公开(公告)日：2009-11-19
申请号：US12414191
申请日：2009-03-30
申请人： Keith Bjornson , Harold Lee , Jonas Korlach , Stephen Turner , Arek Bibillo , Fred Christians , Robin Emig , Molly He , Insil Park , Lei Jia , Andrei Fedorov , John Lyle
发明人： Keith Bjornson , Harold Lee , Jonas Korlach , Stephen Turner , Arek Bibillo , Fred Christians , Robin Emig , Molly He , Insil Park , Lei Jia , Andrei Fedorov , John Lyle
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6869 , C12Q1/6874 , C12Q2527/125 , C12Q2527/119 , C12Q2527/101 , C12Q2565/1015 , C12Q2527/137 , C12Q2521/101
摘要： Compositions, kits, methods and systems for nucleotide sequencing comprising producing polymerase reactions that exhibit two kinetically observable steps within an observable phase of the polymerase reaction. Two slow step systems can be produced, for example, by selecting the appropriate polymerase enzyme, polymerase reaction conditions including cofactors, and polymerase reaction substrates including the primed template and nucleotides.
摘要翻译：用于核苷酸测序的组合物，试剂盒，方法和系统，其包括产生聚合酶反应，其在聚合酶反应的可观察期内显示两个动力学可观察步骤。可以通过例如选择合适的聚合酶，包括辅因子的聚合酶反应条件和包括引发的模板和核苷酸的聚合酶反应底物来产生两个缓慢的步骤系统。

4. 发明授权

US08133672B2 Two slow-step polymerase enzyme systems and methods 有权
标题翻译：两个慢步聚合酶系统和方法
公开(公告)号：US08133672B2
公开(公告)日：2012-03-13
申请号：US12414191
申请日：2009-03-30
申请人： Keith Bjornson , Harold Lee , Jonas Korlach , Stephen Turner , Arek Bibillo , Fred Christians , Robin Emig , Molly He , Insil Park , Lei Jia , Andrei Fedorov , John Lyle
发明人： Keith Bjornson , Harold Lee , Jonas Korlach , Stephen Turner , Arek Bibillo , Fred Christians , Robin Emig , Molly He , Insil Park , Lei Jia , Andrei Fedorov , John Lyle
IPC分类号： C12Q1/68 , C12P19/34 , C07H21/02 , C07H21/04
CPC分类号： C12Q1/6869 , C12Q1/6874 , C12Q2527/125 , C12Q2527/119 , C12Q2527/101 , C12Q2565/1015 , C12Q2527/137 , C12Q2521/101
摘要： Compositions, kits, methods and systems for nucleotide sequencing comprising producing polymerase reactions that exhibit two kinetically observable steps within an observable phase of the polymerase reaction. Two slow step systems can be produced, for example, by selecting the appropriate polymerase enzyme, polymerase reaction conditions including cofactors, and polymerase reaction substrates including the primed template and nucleotides.
摘要翻译：用于核苷酸测序的组合物，试剂盒，方法和系统，其包括产生聚合酶反应，其在聚合酶反应的可观察期内显示两个动力学可观察步骤。可以通过例如选择合适的聚合酶，包括辅因子的聚合酶反应条件和包括引发的模板和核苷酸的聚合酶反应底物来产生两个缓慢的步骤系统。

5. 发明申请

US20120115736A1 NUCLEIC ACID SEQUENCE ANALYSIS 有权
标题翻译：核酸序列分析
公开(公告)号：US20120115736A1
公开(公告)日：2012-05-10
申请号：US13271889
申请日：2011-10-12
申请人： Keith Bjornson , Arek Bibillo , Fred Christians , Kevin Travers , Robin Emig , Stephen Turner
发明人： Keith Bjornson , Arek Bibillo , Fred Christians , Kevin Travers , Robin Emig , Stephen Turner
IPC分类号： C12Q1/68 , C40B20/00
CPC分类号： C12Q1/6869 , C07K14/00 , C12Q1/6874 , C12Q2531/125 , C12Q2521/101
摘要： Provided are methods for sequencing a nucleic acid with a sequencing enzyme, e.g., a polymerase or exonuclease. The sequencing enzyme can optionally be exchanged with a second sequencing enzyme, which continues the sequencing of the nucleic acid. In certain embodiments, a template is fixed to a surface through a template localizing moiety. The template localizing moiety can optionally anneal with the nucleic acid and/or associate with the sequencing enzyme. Also provided are compositions comprising a nucleic acid and a first sequencing enzyme, which can sequence the nucleic acid and optionally exchange with a second sequencing enzyme present in the composition. Compositions in which a template localizing moiety is immobilized on a surface are provided. Also provided are methods for using data from analytical reactions wherein two different enzymes are employed, e.g., at a same or different reaction regions.
摘要翻译：提供了用测序酶（例如聚合酶或外切核酸酶）对核酸进行测序的方法。测序酶可以任选地与第二个测序酶交换，其继续测序核酸。在某些实施方案中，通过模板定位部分将模板固定在表面上。模板定位部分可任选地与核酸退火和/或与测序酶缔合。还提供了包含核酸和第一测序酶的组合物，其可以对核酸进行序列化并且任选与组合物中存在的第二测序酶交换。提供其中将模板定位部分固定在表面上的组合物。还提供了使用来自分析反应的数据的方法，其中使用两种不同的酶，例如在相同或不同的反应区域。

6. 发明授权

US08921046B2 Nucleic acid sequence analysis 有权
标题翻译：核酸序列分析
公开(公告)号：US08921046B2
公开(公告)日：2014-12-30
申请号：US13271889
申请日：2011-10-12
申请人： Keith Bjornson , Arek Bibillo , Fred Christians , Kevin Travers , Robin Emig , Stephen Turner
发明人： Keith Bjornson , Arek Bibillo , Fred Christians , Kevin Travers , Robin Emig , Stephen Turner
IPC分类号： C12Q1/68 , C12P19/34 , C07H21/04
CPC分类号： C12Q1/6869 , C07K14/00 , C12Q1/6874 , C12Q2531/125 , C12Q2521/101
摘要： Provided are methods for sequencing a nucleic acid with a sequencing enzyme, e.g., a polymerase or exonuclease. The sequencing enzyme can optionally be exchanged with a second sequencing enzyme, which continues the sequencing of the nucleic acid. In certain embodiments, a template is fixed to a surface through a template localizing moiety. The template localizing moiety can optionally anneal with the nucleic acid and/or associate with the sequencing enzyme. Also provided are compositions comprising a nucleic acid and a first sequencing enzyme, which can sequence the nucleic acid and optionally exchange with a second sequencing enzyme present in the composition. Compositions in which a template localizing moiety is immobilized on a surface are provided. Also provided are methods for using data from analytical reactions wherein two different enzymes are employed, e.g., at a same or different reaction regions.
摘要翻译：提供了用测序酶（例如聚合酶或外切核酸酶）对核酸进行测序的方法。测序酶可以任选地与第二个测序酶交换，其继续测序核酸。在某些实施方案中，通过模板定位部分将模板固定在表面上。模板定位部分可任选地与核酸退火和/或与测序酶缔合。还提供了包含核酸和第一测序酶的组合物，其可以对核酸进行序列化并且任选与组合物中存在的第二测序酶交换。提供其中将模板定位部分固定在表面上的组合物。还提供了使用来自分析反应的数据的方法，其中使用两种不同的酶，例如在相同或不同的反应区域。

7. 发明申请

US20090176233A1 Screening assays for polymerase enhancement 审中-公开
标题翻译：聚合酶增强筛选试验
公开(公告)号：US20090176233A1
公开(公告)日：2009-07-09
申请号：US12315844
申请日：2008-12-05
申请人： Sonya Clark , Homero L. Rey , Fred Christians , Jonas Korlach
发明人： Sonya Clark , Homero L. Rey , Fred Christians , Jonas Korlach
IPC分类号： C12Q1/68 , C12N9/00 , C12M1/00
CPC分类号： C12N9/1252 , C12Q1/48 , G01N2333/9126
摘要： Methods of screening for and selecting for improved polymerases suitable for single molecule sequencing are provided.
摘要翻译：提供了筛选和选择适用于单分子测序的改良聚合酶的方法。

8. 发明授权

US08257954B2 Generation of modified polymerases for improved accuracy in single molecule sequencing 有权
标题翻译：修饰聚合酶的生成，提高了单分子测序的准确性
公开(公告)号：US08257954B2
公开(公告)日：2012-09-04
申请号：US12384112
申请日：2009-03-30
申请人： Sonya Clark , Arek Bibillo , Paul Peluso , Fred Christians , Molly He , Insil Park , Harold Lee , Keith Bjornson , Lei Jia , Robin Emig
发明人： Sonya Clark , Arek Bibillo , Paul Peluso , Fred Christians , Molly He , Insil Park , Harold Lee , Keith Bjornson , Lei Jia , Robin Emig
IPC分类号： C12N9/12 , C12P19/34
CPC分类号： C12N9/1252 , C12Q1/6844 , C12Q1/6869 , C12Q2521/101
摘要： Provided are compositions comprising modified recombinant polymerases that exhibit branching fractions that are less than the branching fractions of the polymerases from which they were derived, or branching fractions that are less than about 25% for a phosphate-labeled nucleotide analog. Also provided are compositions comprising modified recombinant polymerases that exhibit closed polymerase/DNA complexes with increased stability relative to the parental polymerases. Also provided are compositions comprising modified recombinant polymerases that exhibit decreased rate constants relative to the parental polymerases. Provided are methods for generating polymerases with the aforementioned phenotypes. Provided are methods of using such polymerases to make a DNA or to sequence a DNA template.
摘要翻译：提供了包含修饰的重组聚合酶的组合物，其表现出小于其衍生自其的聚合酶的支化部分的支化部分，或对于磷酸酯标记的核苷酸类似物的支化部分小于约25％。还提供了包含修饰的重组聚合酶的组合物，其表现出相对于亲本聚合酶具有增加的稳定性的闭合的聚合酶/ DNA复合物。还提供了包含修饰的重组聚合酶的组合物，其相对于亲本聚合酶表现出降低的速率常数。提供了产生具有上述表型的聚合酶的方法。提供使用这种聚合酶制备DNA或顺序DNA模板的方法。

9. 发明申请

US20100075328A1 IMMOBILIZED NUCLEIC ACID COMPLEXES FOR SEQUENCE ANALYSIS 有权
标题翻译：用于序列分析的固定核酸复合物
公开(公告)号：US20100075328A1
公开(公告)日：2010-03-25
申请号：US12562690
申请日：2009-09-18
申请人： KEITH BJORNSON , Arek Bibillo , Fred Christians , Kevin Travers , Robin Emig
发明人： KEITH BJORNSON , Arek Bibillo , Fred Christians , Kevin Travers , Robin Emig
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6874 , C12Q2531/125
摘要： Provided are methods for sequencing a nucleic acid that include fixing a template to a surface through a template localizing moiety and sequencing the nucleic acid with a sequencing enzyme, e.g. a polymerase or exonuclease. The sequencing enzyme can optionally be exchanged with a second sequencing enzyme, which continues the sequencing of the nucleic acid. The template localizing moiety can optionally anneal with the nucleic acid and/or associate with the sequencing enzyme. Also provided are compositions comprising a nucleic acid fixed to a surface via a template localizing moiety, and a first sequencing enzyme, which can sequence the nucleic acid and optionally exchange with a second sequencing enzyme present in the composition. Compositions in which a template localizing moiety is immobilized on a surface are provided. Compositions for sequencing reactions are provided. Also provided are sequencing systems comprising reaction regions in which or near which template localizing moieties are immobilized.
摘要翻译：提供了用于对核酸进行测序的方法，其包括通过模板定位部分将模板固定到表面，并用测序酶（例如）测序核酸。聚合酶或外切核酸酶。测序酶可以任选地与第二个测序酶交换，其继续测序核酸。模板定位部分可任选地与核酸退火和/或与测序酶缔合。还提供了包含通过模板定位部分固定在表面上的核酸的组合物和可以使序列核酸并任选地与组合物中存在的第二测序酶交换的第一测序酶。提供其中将模板定位部分固定在表面上的组合物。提供了测序反应的组合物。还提供了包含反应区的测序系统，其中固定有定位部分的模板定位部分。

10. 发明申请

US20110189659A1 Generation of modified polymerases for improved accuracy in single molecule sequencing 有权
标题翻译：修饰聚合酶的生成，提高了单分子测序的准确性
公开(公告)号：US20110189659A1
公开(公告)日：2011-08-04
申请号：US12924701
申请日：2010-09-30
申请人： Sonya Clark , Arek Bibillo , Paul Peluso , Fred Christians , Molly He , Insil Park , Harold Lee , Keith Bjornson , Lei Jia , Robin Emig
发明人： Sonya Clark , Arek Bibillo , Paul Peluso , Fred Christians , Molly He , Insil Park , Harold Lee , Keith Bjornson , Lei Jia , Robin Emig
IPC分类号： C12N9/12 , C12Q1/68 , C12N11/00 , C12P19/34
CPC分类号： C12N9/12 , C12N11/00 , C12P19/34 , C12Q1/68
摘要： Provided are compositions comprising recombinant DNA polymerases that include amino acid substitutions, insertions, deletions and/or heterologous or exogenous features that confer modified properties upon the polymerase for enhanced single molecule sequencing. Such properties can include reduced reaction rates at one or more steps of the polymerase kinetic cycle, increased closed polymerase/DNA complex stability, enhanced metal ion coordination, reduced exonuclease activity, decreased branching fractions, and the like. Polymerases that exhibit branching fractions that are less than the branching fractions of the polymerases from which they were derived, or branching fractions that are less than about 25% for a phosphate-labeled nucleotide analog, are also provided. Also provided are nucleic acids which encode the polymerases with the aforementioned phenotypes, as well as methods of using such polymerases to make a DNA or to sequence a DNA template.
摘要翻译：提供了包含重组DNA聚合酶的组合物，其包括氨基酸取代，插入，缺失和/或异源或外源特征，其在聚合酶上赋予改进的性质以用于增强的单分子测序。这种性质可以包括聚合酶动力学循环的一个或多个步骤的降低的反应速率，增加的闭合聚合酶/ DNA复合物稳定性，增强的金属离子配位，减少的核酸外切酶活性，降低的支化部分等。还提供了表现出低于其衍生的聚合酶的支化级分的支化级分的聚合酶，或者对于磷酸酯标记的核苷酸类似物的支化级分小于约25％。还提供了编码具有上述表型的聚合酶的核酸，以及使用这种聚合酶制备DNA或顺序DNA模板的方法。

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