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21. 发明授权

US10207240B2 Methods and microfluidic devices for the manipulation of droplets in high fidelity polynucleotide assembly 有权
公开(公告)号：US10207240B2
公开(公告)日：2019-02-19
申请号：US13505646
申请日：2010-11-03
申请人： Joseph Jacobson , Larry Li-Yang Chu , Senthil Ramu
发明人： Joseph Jacobson , Larry Li-Yang Chu , Senthil Ramu
IPC分类号： C40B60/14 , C40B60/00 , C40B40/06 , B01J19/00 , B01L3/00 , B82Y30/00 , C12Q1/68 , B01L3/02 , B01L7/00
摘要： Methods and devices are provided for manipulating droplets on a support using surface tension properties, moving the droplets along a predetermined path and merging two droplets together enabling a number of chemical reactions. Disclosed are methods for controlling the droplets volumes. Disclosed are methods and devices for synthesizing at least one oligonucleotide having a predefined sequence. Disclosed are methods and devices for synthesizing and/or assembling at least one polynucleotide product having a predefined sequence from a plurality of different oligonucleotides having a predefined sequence. In exemplary embodiments, the methods involve synthesis and/or amplification of different oligonucleotides immobilized on a solid support, release of synthesized/amplified oligonucleotides in solution to form droplets, recognition and removal of error-containing oligonucleotides, moving or combining two droplets to allow hybridization and/or ligation between two different oligonucleotides, and further chain extension reaction following hybridization and/or ligation to hierarchically generate desired length of polynucleotide products.

22. 发明授权

US10053719B2 Compositions and methods for synthesis of high fidelity oligonucleotides 有权
公开(公告)号：US10053719B2
公开(公告)日：2018-08-21
申请号：US14765978
申请日：2014-03-13
申请人： Gen9, Inc.
发明人： Ishtiaq E. Saaem , Michael E. Hudson , Stephen F. Archer , Sophea Sann-Rorth , Ellen J. Sanchez , Joseph Jacobson
IPC分类号： C12P19/34 , C12N15/10 , C12N15/66
CPC分类号： C12P19/34 , C12N15/10 , C12N15/1031 , C12N15/66
摘要： Aspects of the invention relate to methods, compositions for synthesizing high fidelity oligonucleotides.

23. 发明授权

US09150853B2 Methods for screening proteins using DNA encoded chemical libraries as templates for enzyme catalysis 有权
标题翻译：使用DNA编码化学文库筛选蛋白质作为酶催化模板的方法
公开(公告)号：US09150853B2
公开(公告)日：2015-10-06
申请号：US13799039
申请日：2013-03-13
申请人： Gen9, Inc.
发明人： Michael E. Hudson , Joseph Jacobson
IPC分类号： C12N15/10
CPC分类号： C12N15/1034 , C12N15/1037 , C12N15/1086
摘要： Disclosed are methods, compositions and devices for screening a protein library for proteins having a desired activity, such as capable of catalyzing the formation of a bond between two reactants. In an exemplary embodiments, a plurality of proteins are expressed in vitro from a plurality of nucleic acids, the plurality of proteins are exposed with two single stranded oligonucleotides having complementary sequences, each oligonucleotide having a reactant and a fluorophore, the fluorescence of the protein-reactant-oligonucleotide-fluorophore complexes is detected and the complexes showing detectable fluorescence energy transfer are isolated, thereby isolating proteins having the desired enzymatic activity.
摘要翻译：公开了用于筛选具有所需活性的蛋白质的蛋白质文库的方法，组合物和装置，例如能够催化两种反应物之间的键的形成。在一个示例性实施方案中，多个蛋白质在体外从多个核酸表达，多个蛋白质用两个具有互补序列的单链寡核苷酸暴露，每个寡核苷酸具有反应物和荧光团，蛋白质 - 检测到反应物 - 寡核苷酸 - 荧光团复合物，并分离出显示可检测的荧光能量转移的复合物，从而分离具有所需酶活性的蛋白质。

24. 发明授权

US09051666B2 Microarray synthesis and assembly of gene-length polynucleotides 有权
标题翻译：基因长度多核苷酸的微阵列合成和装配
公开(公告)号：US09051666B2
公开(公告)日：2015-06-09
申请号：US13617685
申请日：2012-09-14
申请人： Andrew V. Oleinikov
发明人： Andrew V. Oleinikov
IPC分类号： C12Q1/68 , C12P19/34 , C40B50/14 , B01J19/00 , B82Y30/00 , C40B80/00 , C40B40/06
CPC分类号： C12N15/1068 , B01J19/0046 , B01J2219/00378 , B01J2219/00385 , B01J2219/00432 , B01J2219/00454 , B01J2219/005 , B01J2219/00527 , B01J2219/00585 , B01J2219/00596 , B01J2219/00605 , B01J2219/00608 , B01J2219/00626 , B01J2219/00639 , B01J2219/00641 , B01J2219/00659 , B01J2219/00675 , B01J2219/00677 , B01J2219/00689 , B01J2219/00713 , B01J2219/00722 , B82Y30/00 , C12Q1/6837 , C40B40/06 , C40B50/14 , C40B80/00
摘要： There is disclosed a process for in vitro synthesis and assembly of long, gene-length polynucleotides based upon assembly of multiple shorter oligonucleotides synthesized in situ on a microarray platform. Specifically, there is disclosed a process for in situ synthesis of oligonucleotide fragments on a solid phase microarray platform and subsequent, “on device” assembly of larger polynucleotides composed of a plurality of shorter oligonucleotide fragments.
摘要翻译：公开了一种基于在微阵列平台上原位合成的多个较短的寡核苷酸的组装的体外合成和装配长的基因长度多核苷酸的方法。具体地，公开了在固相微阵列平台上原位合成寡核苷酸片段的方法，随后在由多个较短的寡核苷酸片段组成的较大多核苷酸的“装置上”组装上。

25. 发明授权

US08808986B2 Methods and devices for high fidelity polynucleotide synthesis 有权
标题翻译：用于高保真多核苷酸合成的方法和装置
公开(公告)号：US08808986B2
公开(公告)日：2014-08-19
申请号：US13060178
申请日：2009-08-27
申请人： Joseph Jacobson , George Church , Larry Li-Yang Chu
发明人： Joseph Jacobson , George Church , Larry Li-Yang Chu
IPC分类号： C12Q1/68 , C12P19/34 , C07H21/00 , C07H21/02 , C07H21/04
CPC分类号： C12N15/1093 , B01J2219/00378 , B01J2219/00389 , B01J2219/0052 , B01J2219/00529 , B01J2219/00585 , B01J2219/0059 , B01J2219/00596 , B01J2219/00599 , B01J2219/00608 , B01J2219/00659 , B01J2219/00693 , B01J2219/00722 , B01L3/0268 , B01L3/502792 , B01L2400/0436 , B01L2400/0439 , C12N15/10 , C12N15/66 , C12Q1/6837 , C12Q1/6844 , C12Q2525/131 , C12Q2521/514 , C12Q2565/501
摘要： Disclosed are methods for synthesizing and/or assembling at least one polynucleotide product having a predefined sequence from a plurality of different oligonucleotides. In exemplary embodiments, the methods involve synthesis and/or amplification of different oligonucleotides immobilized on a solid support, release of synthesized/amplified oligonucleotides in solution to form droplets, recognition and removal of error-containing oligonucleotides, moving or combining two droplets to allow hybridization and/or ligation between two different oligonucleotides, and further chain extension reaction following hybridization and/or ligation to hierarchically generate desired length of polynucleotide products.
摘要翻译：公开了用于从多种不同寡核苷酸合成和/或组装具有预定序列的至少一种多核苷酸产物的方法。在示例性实施方案中，所述方法涉及固定在固体支持物上的不同寡核苷酸的合成和/或扩增，在溶液中释放合成/扩增的寡核苷酸以形成液滴，识别和去除含错误的寡核苷酸，移动或组合两个液滴以允许杂交和/或两个不同寡核苷酸之间的连接，以及杂交和/或连接之后的进一步的链延伸反应以分级地产生所需长度的多核苷酸产物。

26. 发明申请

US20120028843A1 Methods and Apparatuses for Chip-Based DNA Error Reduction 有权
标题翻译：基于芯片的DNA错误减少的方法和设备
公开(公告)号：US20120028843A1
公开(公告)日：2012-02-02
申请号：US13164045
申请日：2011-06-20
申请人： Senthil Ramu , Joseph Jacobson
发明人： Senthil Ramu , Joseph Jacobson
IPC分类号： C40B50/06 , C07H21/00 , C12P19/34
CPC分类号： C12N15/1058 , C12Q1/6848 , C12Q2521/514 , C12Q2565/537
摘要： Methods and apparatus relate to reduction of sequence errors generated during synthesis of nucleic acids on a microarray chip. The error reduction can include synthesis of complementary stands (to template strands), using a short universal primer complementary to the template strands and polymerase. Heteroduplex can be formed be melting and re-annealing complementary stands and template strands. The heteroduplexes containing a mismatch can be recognized and cleaved by a mismatch endonuclease. The mismatch-containing cleaved heteroduplexes can be removed from the microarray chip using a global buffer exchange. The error free synthetic nucleic acids generated therefrom can be used for a variety of applications, including synthesis of biofuels and value-added pharmaceutical products.
摘要翻译：方法和装置涉及减少在微阵列芯片上的核酸合成期间产生的序列错误。错误减少可以包括使用与模板链和聚合酶互补的短通用引物来合成互补基团（模板链）。 Heteroduplex可以形成为熔化和重新退火互补架和模板链。含有不匹配的异源双链可以被不匹配的内切核酸酶识别和切割。可以使用全局缓冲液交换从微阵列芯片去除包含错配的切割的异源双链体。由此产生的无错误的合成核酸可用于各种应用，包括合成生物燃料和增值药物产品。

27. 发明授权

US11845054B2 Methods and devices for nucleic acids synthesis 有权
公开(公告)号：US11845054B2
公开(公告)日：2023-12-19
申请号：US17378133
申请日：2021-07-16
申请人： Gen9, Inc.
发明人： Joseph Jacobson , Li-Yun A. Kung , Andrew Kirk Wilson , Senthil Ramu , Daniel Schindler , Michael E. Hudson
IPC分类号： C12Q1/68 , B01J19/00 , C12N15/10 , C12N15/66 , C12P19/34
CPC分类号： B01J19/0046 , C12N15/10 , C12N15/1031 , C12N15/66 , B01J2219/00585 , B01J2219/00596 , B01J2219/00722 , C12P19/34 , C12N15/10 , C12Q2521/343 , C12Q2521/50 , C12N15/10 , C12Q2521/50 , C12Q2521/501 , C12N15/10 , C12N15/64
摘要： Methods and apparatus relate to the synthesis of polynucleotides having a predefined sequence on a support. Assembly methods include primer extension to generate overlapping construction oligonucleotides and assembly of the polynucleotides of interest onto an anchor support-bound oligonucleotides. Methods and apparatus for selection of polynucleotides having the predefined sequence and/or length are disclosed.

28. 发明授权

US10982208B2 Protein arrays and methods of using and making the same 有权
公开(公告)号：US10982208B2
公开(公告)日：2021-04-20
申请号：US16578488
申请日：2019-09-23
申请人： Gen9, Inc.
发明人： Joseph Jacobson , Michael E. Hudson , Larry Li-Yang Chu
IPC分类号： C12N15/10 , B01J19/00 , C40B50/14
摘要： Methods and devices are provided for preparing a protein array having a plurality of proteins. In one embodiment, the method includes providing a plurality of nucleic acids each having a predefined sequence and expressing in vitro a plurality of proteins from the plurality of nucleic acids. In another embodiment, protein arrays having a solid surface and a microvolume are also provided. The solid surface can have a plurality of anchor oligonucleotides capable of hybridizing with a plurality of nucleic acids. The microvolume can cover each of the plurality of anchor oligonucleotides and can be configured to produce a polypeptide from each of the plurality of nucleic acids.

29. 发明申请

US20210062185A1 MICROARRAY SYNTHESIS AND ASSEMBLY OF GENE-LENGTH POLYNUCLEOTIDES 有权
公开(公告)号：US20210062185A1
公开(公告)日：2021-03-04
申请号：US17019448
申请日：2020-09-14
申请人： Gen9, Inc.
发明人： Andrew V. Oleinikov
IPC分类号： C12N15/10 , B82Y30/00 , C40B80/00 , B01J19/00 , C40B50/14 , C12Q1/6837
摘要： There is disclosed a process for in vitro synthesis and assembly of long, gene-length polynucleotides based upon assembly of multiple shorter oligonucleotides synthesized in situ on a microarray platform. Specifically, there is disclosed a process for in situ synthesis of oligonucleotide fragments on a solid phase microarray platform and subsequent, “on device” assembly of larger polynucleotides composed of a plurality of shorter oligonucleotide fragments.

30. 发明授权

US10927369B2 Methods for sorting nucleic acids and multiplexed preparative in vitro cloning 有权
公开(公告)号：US10927369B2
公开(公告)日：2021-02-23
申请号：US16039288
申请日：2018-07-18
申请人： Gen9, Inc.
发明人： Joseph Jacobson , Martin J. Goldberg , Li-Yun A. Kung , Daniel Schindler , Michael E. Hudson
IPC分类号： C12N15/10
摘要： Methods and compositions relate to the sorting and cloning of high fidelity nucleic acids using high throughput sequencing. Specifically, nucleic acid molecules having the desired predetermined sequence can be sorted from a pool comprising a plurality of nucleic acids having correct and incorrect sequences.

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