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公开(公告)号：US20220364168A1
公开(公告)日：2022-11-17
申请号：US17719362
申请日：2022-04-12
Applicant: Pacific Biosciences of California, Inc.
Inventor： Jaime Juan Benitez-Marzan , Natasha Popovich , Lei Sun , Satwik Kamtekar , Keith Bjornson , Jeremiah Hanes , Karl Voss , Erik Miller , Thomas Linsky , Leewin Chern
IPC: C12Q1/6874 , B01J19/00
Abstract: The present invention provides methods, compositions, and systems for distributing molecules and complexes into reaction sites. In particular, the methods, compositions, and systems of the present invention result in loading of polymerase enzyme complexes into a predetermined number of reaction sites, including nanoscale wells.
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公开(公告)号：US10787573B2
公开(公告)日：2020-09-29
申请号：US16209916
申请日：2018-12-04
Applicant: Pacific Biosciences of California, Inc.
Inventor： Frank Zheng , Jeremiah Hanes , Gene Shen , Louis Brogley , Stephen Yue , Yuri Lapin , John Lyle , Honey Osuna , Andrei Fedorov , Lubomir Sebo
IPC: C09B69/10 , C09B23/10 , C09B23/06 , C09B69/00 , G01N33/533
Abstract: Multimeric protected fluorescent reagents and their methods of synthesis are provided. The reagents are useful in various fluorescence-based analytical methods, including the analysis of highly multiplexed optical reactions in large numbers at high densities, such as single molecule real time nucleic acid sequencing reactions. The reagents contain fluorescent dye elements, that allow the compounds to be detected with high sensitivity at desirable wavelengths, binding elements, that allow the compounds to be recognized specifically by target biomolecules, and protective shield elements, that decrease undesirable contacts between the fluorescent dye elements and the bound target biomolecules and that therefore decrease photodamage of the bound target biomolecules by the fluorescent dye elements. The reagents also contain coupling elements connect monomeric compounds into multimeric forms, thereby increasing brightness.
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公开(公告)号：US20180023134A1
公开(公告)日：2018-01-25
申请号：US15656622
申请日：2017-07-21
Applicant: Pacific Biosciences of California, Inc.
Inventor： Jeremiah Hanes , Satwik Kamtekar , Yufeng Qian , Keith Bjornson , Thang Pham , David Rank , Paul Peluso , Erik Miller , Kristin Gleitsman , Leewin Chern , Steven Lin , Stephen Turner
IPC: C12Q1/68 , G01N27/447 , G01N33/487
Abstract: The present invention provides methods, compositions, and systems for distributing molecules and complexes into reaction sites. In particular, the methods, compositions, and systems of the present invention result in an active loading of molecules and complexes into reaction sites with improved efficiency over loading by passive diffusion methods alone.
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公开(公告)号：US09678080B2
公开(公告)日：2017-06-13
申请号：US14303296
申请日：2014-06-12
Applicant: Pacific Biosciences of California, Inc.
Inventor： Erik Miller , Satwik Kamtekar , Keith Bjornson , Jeremiah Hanes
Abstract: Multi-biotinylated reactants are provided which can be used in divalent complexes for various applications such as colocalization, labeling, immobilization, and purification. Methods for constructing, purifying, and using the bis-biotinylated reactants are also provided. In certain embodiments, two bis-biotinylated reactants are bound to a single streptavidin tetramer to provide a complex having a 1:1 stoichiometry with respect to the bis-biotinylated reactants.
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公开(公告)号：US20170037462A1
公开(公告)日：2017-02-09
申请号：US15227661
申请日：2016-08-03
Applicant: Pacific Biosciences of California, Inc.
Inventor： Stephen Turner , Jonas Korlach , Satwik Kamtekar , Jeremiah Hanes
IPC: C12Q1/68
CPC classification number: C12Q1/6869 , B82Y10/00 , B82Y15/00 , G01N27/4145 , G01N27/4146 , G01N27/4148 , G11C19/28 , H01L29/0669 , H01L29/0673 , H01L29/1606 , C12Q2533/101 , C12Q2565/607
Abstract: Real time electronic sequencing methods, devices, and systems are described. Arrays of nanoFET devices are used to provide sequence information about a template nucleic acid in a polymerase-template complex bound to the nanoFET. A sequencing reaction mixture comprising nucleotide analogs having conductivity labels is introduced to the array of nanoscale electronic elements comprising nanoFETs under conditions of polymerase mediated nucleic acid synthesis. The polymerase enzyme template complex is attached to the gate of the nanoFET in an orientation whereby the nucleotide exit region of the polymerase enzyme is directed toward the gate of the nanoFET. Methods for producing nanoFET arrays are provided.
Abstract translation: 描述了实时电子测序方法，设备和系统。 纳米器件阵列用于提供与纳米晶体结合的聚合酶 - 模板复合物中关于模板核酸的序列信息。 包含具有电导率标记的核苷酸类似物的测序反应混合物在聚合酶介导的核酸合成的条件下被引入到纳米级电子元件阵列中，该纳米级电子元件包括纳米晶体。 聚合酶模板复合物以定向方式连接到纳米器件的栅极，由此聚合酶的核苷酸出口区域指向纳米器件的栅极。 提供了制造nanoFET阵列的方法。
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公开(公告)号：US20150330987A1
公开(公告)日：2015-11-19
申请号：US14727553
申请日：2015-06-01
Applicant: Pacific Biosciences of California, Inc.
Inventor： Keith Bjornson , Jeremiah Hanes , Erik Miller , Satwik Kamtekar , Lubomir Sebo , Louis Brogley
CPC classification number: C07H21/04 , C07H19/06 , C07H19/16 , C07K14/36 , C12N9/10 , C12Q1/6869 , C12Q1/6874 , C12Q2523/313 , C12Q2535/113 , C12Q2563/107 , C12Q2563/131 , G01N33/582 , G01N33/583
Abstract: Compositions, methods, and systems are provided for nucleotide analogs comprising protein shields for improving enzyme photostability in single molecule real time sequencing. Nucleotide analogs of the invention have a protein shield between the dye moieties and nucleotide moieties of the analog. The nucleotide analogs have two avidin proteins connected by a central dye component, and each avidin protein is further connected to a nucleotide component. The protein can prevent the direct interaction of the dye moiety with the enzyme carrying out nucleotide synthesis preventing photodamage to the enzyme. The nucleotide analogs of the invention can have multiple dyes and multiple nucleotide moieties.
Abstract translation: 提供了包含用于提高单分子实时测序中酶光稳定性的蛋白质屏蔽物的核苷酸类似物的组合物，方法和系统。 本发明的核苷酸类似物在染料部分和类似物的核苷酸部分之间具有蛋白质屏蔽。 核苷酸类似物具有通过中心染料组分连接的两个亲和素蛋白，并且每个抗生物素蛋白蛋白进一步连接到核苷酸组分。 该蛋白质可以防止染料部分与进行核苷酸合成的酶直接相互作用，从而防止对酶的光损伤。 本发明的核苷酸类似物可以具有多个染料和多个核苷酸部分。
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公开(公告)号：US20150203908A1
公开(公告)日：2015-07-23
申请号：US14642307
申请日：2015-03-09
Applicant: Pacific Biosciences of California, Inc.
Inventor： Andrei Fedorov , John Lyle , Keith Bjornson , Jeremiah Hanes
IPC: C12Q1/68
CPC classification number: C12Q1/6869 , C12Q2527/125 , C12Q2563/137
Abstract: Compositions, kits, methods and systems for single molecule nucleotide sequencing comprising producing polymerase reactions having lithium that control the median pulse width for incorporated nucleotides are disclosed. The levels of lithium are used to control pulse width while allowing other sequencing parameters to remain within a desirable range.
Abstract translation: 公开了用于单分子核苷酸测序的组合物，试剂盒，方法和系统，其包括产生具有控制掺入的核苷酸的中值脉冲宽度的具有锂的聚合酶反应。 锂的水平用于控制脉冲宽度，同时允许其他测序参数保持在期望的范围内。
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公开(公告)号：US20130316912A1
公开(公告)日：2013-11-28
申请号：US13767619
申请日：2013-02-14
Applicant: Pacific Biosciences of California, Inc.
Inventor： Keith Bjornson , Jeremiah Hanes , Erik Miller , Satwik Kamtekar , Lubomir Sebo , Louis Brogley
CPC classification number: C07H21/04 , C07H19/06 , C07H19/16 , C07K14/36 , C12N9/10 , C12Q1/6869 , C12Q1/6874 , C12Q2523/313 , C12Q2535/113 , C12Q2563/107 , C12Q2563/131 , G01N33/582 , G01N33/583
Abstract: Compositions and methods are provided for nucleotide analogs comprising protein shields for improving enzyme photostability in single molecule real time sequencing. Nucleotide analogs of the invention have a protein shield between the dye moieties and nucleotide moieties of the analog. The protein prevents the direct interaction of the dye moiety with the enzyme carrying out nucleotide synthesis preventing photodamage to the enzyme. The nucleotide analogs of the invention can have multiple dyes and multiple nucleotide moieties.
Abstract translation: 提供了包含用于提高单分子实时测序中酶光稳定性的蛋白质屏蔽物的核苷酸类似物的组合物和方法。 本发明的核苷酸类似物在染料部分和类似物的核苷酸部分之间具有蛋白质屏蔽。 蛋白质可以防止染料部分与进行核苷酸合成的酶直接相互作用，从而防止酶对光的损伤。 本发明的核苷酸类似物可以具有多个染料和多个核苷酸部分。
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公开(公告)号：US20250002878A1
公开(公告)日：2025-01-02
申请号：US18665785
申请日：2024-05-16
Applicant: Pacific Biosciences of California, Inc.
Inventor： Satwik Kamtekar , Arek Bibillo , Keith Bjornson , Fred Christians , Colleen Cutcliffe , Jeremiah Hanes , Lei Jia , Walter Lee , Erik Miller , Pranav Patel
IPC: C12N9/12 , C12P19/34 , C12Q1/6869
Abstract: Provided are compositions comprising recombinant DNA polymerases that include amino acid substitutions, insertions, deletions, and/or exogenous features that confer modified properties upon the polymerase for enhanced single molecule sequencing. Such properties include increased resistance to photodamage, and can also include enhanced metal ion coordination, reduced exonuclease activity, reduced reaction rates at one or more steps of the polymerase kinetic cycle, decreased branching fraction, altered cofactor selectivity, increased yield, increased thermostability, increased accuracy, increased speed, increased readlength, and the like. 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.
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公开(公告)号：US11781177B2
公开(公告)日：2023-10-10
申请号：US16841450
申请日：2020-04-06
Applicant: Pacific Biosciences of California, Inc.
Inventor： Satwik Kamtekar , Lubomir Sebo , Leewin Chern , Thomas Linsky , Jeremiah Hanes , Erik Miller , Ying Yang , Stephen Yue
IPC: C07H21/02 , C12Q1/6848 , C12N9/22 , C07K14/36
CPC classification number: C12Q1/6848 , C07K14/36 , C12N9/22 , C12Q2535/00 , C12Q2537/143
Abstract: Compositions comprising covalently modified and mutated biotin-binding proteins, particularly biotin-binding proteins having a negative charge at physiological pH, are provided. Methods of producing such proteins are also provided, as are methods of immobilizing, sequencing, and making nucleic acids employing such proteins.
