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1. 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.

2. 发明申请

US20120220497A1 Methods and Microfluidic Devices for the Manipulation of Droplets in High Fidelity Polynucleotide Assembly 审中-公开
标题翻译：用于在高保真多核苷酸装配中操纵水滴的方法和微流体装置
公开(公告)号：US20120220497A1
公开(公告)日：2012-08-30
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： C40B50/14 , C07H1/00 , C40B40/06
CPC分类号： B01J19/0046 , B01J2219/00378 , B01J2219/00495 , B01J2219/00608 , B01J2219/00621 , B01J2219/00644 , B01J2219/0065 , B01J2219/00653 , B01J2219/00659 , B01J2219/00675 , B01J2219/00722 , B01L3/0262 , B01L3/0268 , B01L3/502792 , B01L3/5088 , B01L7/52 , B01L2300/10 , B01L2300/1861 , B01L2400/0427 , B01L2400/043 , B01L2400/0439 , B01L2400/0454 , B82Y30/00 , C12Q2565/629
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.
摘要翻译：提供的方法和装置用于使用表面张力特性来操纵支撑体上的液滴，沿着预定路径移动液滴并将两个液滴合并在一起，从而实现多个化学反应。公开了用于控制液滴体积的方法。公开了用于合成具有预定序列的至少一种寡核苷酸的方法和装置。公开了用于从具有预定序列的多种不同寡核苷酸合成和/或组装至少一种具有预定序列的多核苷酸产物的方法和装置。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。

3. 发明申请

US20130309725A1 Methods and Devices for In Situ Nucleic Acid Synthesis 有权
标题翻译：用于原位核酸合成的方法和装置
公开(公告)号：US20130309725A1
公开(公告)日：2013-11-21
申请号：US13700291
申请日：2011-05-26
申请人： Joseph Jacobson , Senthil Ramu , Daniel Schindler
发明人： Joseph Jacobson , Senthil Ramu , Daniel Schindler
IPC分类号： C12P19/34
CPC分类号： C12N15/1068 , C12N15/1093 , C12N15/66 , C12P19/34
摘要： Disclosed are compositions, methods and devices for the in situ synthesis of nucleic acids. In an exemplary embodiment, a support-bound oligonucleotide is elongated by addition of one or more nucleotides by hybridization of a partially double-stranded oligonucleotide, ligation and removal of unwanted nucleotides.
摘要翻译：公开了用于原位合成核酸的组合物，方法和装置。在一个示例性实施方案中，通过部分双链寡核苷酸的杂交，连接和去除不需要的核苷酸，加入一个或多个核苷酸来延伸载体结合的寡核苷酸。

4. 发明授权

US09422600B2 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：US09422600B2
公开(公告)日：2016-08-23
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： C12P19/34 , C12Q1/68 , C12M1/34 , C12M3/00 , C07H21/02 , C07H21/04 , C07H21/00
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。

5. 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。

6. 发明授权

US09295965B2 Methods and devices for nucleic acid synthesis 有权
标题翻译：用于核酸合成的方法和装置
公开(公告)号：US09295965B2
公开(公告)日：2016-03-29
申请号：US13884463
申请日：2011-11-10
申请人： Joseph Jacobson , Li-yun A. Kung , Andrew Kirk Wilson , Senthil Ramu , Daniel Schindler , Mike Hudson
发明人： Joseph Jacobson , Li-yun A. Kung , Andrew Kirk Wilson , Senthil Ramu , Daniel Schindler , Mike Hudson
IPC分类号： C12Q1/68 , B01J19/00 , C12N15/10 , C12N15/66 , C12P19/34
CPC分类号： B01J19/0046 , B01J2219/00585 , B01J2219/00596 , B01J2219/00722 , C12N15/10 , C12N15/1031 , C12N15/66 , C12P19/34 , C12Q2521/343 , C12Q2521/50 , C12Q2521/501 , 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.
摘要翻译：方法和装置涉及在载体上具有预定序列的多核苷酸的合成。组装方法包括产生重叠构建寡核苷酸的引物延伸和目标多核苷酸组装到锚定载体结合的寡核苷酸上。公开了用于选择具有预定义的序列和/或长度的多核苷酸的方法和装置。

7. 发明授权

US09187777B2 Methods and devices for in situ nucleic acid synthesis 有权
标题翻译：用于原位核酸合成的方法和装置
公开(公告)号：US09187777B2
公开(公告)日：2015-11-17
申请号：US13700291
申请日：2011-05-26
申请人： Joseph Jacobson , Senthil Ramu , Daniel Schindler
发明人： Joseph Jacobson , Senthil Ramu , Daniel Schindler
IPC分类号： C12Q1/68 , C12P19/34 , C12N15/10 , C12N15/66
CPC分类号： C12N15/1068 , C12N15/1093 , C12N15/66 , C12P19/34
摘要： Disclosed are compositions, methods and devices for the in situ synthesis of nucleic acids. In an exemplary embodiment, a support-bound oligonucleotide is elongated by addition of one or more nucleotides by hybridization of a partially double-stranded oligonucleotide, ligation and removal of unwanted nucleotides.
摘要翻译：公开了用于原位合成核酸的组合物，方法和装置。在一个示例性实施方案中，通过部分双链寡核苷酸的杂交，连接和去除不需要的核苷酸，加入一个或多个核苷酸来延伸载体结合的寡核苷酸。

8. 发明申请

US20130296194A1 Methods and Devices for Nucleic Acid Synthesis 有权
标题翻译：核酸合成的方法和装置
公开(公告)号：US20130296194A1
公开(公告)日：2013-11-07
申请号：US13884463
申请日：2011-11-10
申请人： Joseph Jacobson , Li-yun A. Kung , Andrew Kirk Wilson , Senthil Ramu , Daniel Schindler , Mike Hudson
发明人： Joseph Jacobson , Li-yun A. Kung , Andrew Kirk Wilson , Senthil Ramu , Daniel Schindler , Mike Hudson
IPC分类号： B01J19/00
CPC分类号： B01J19/0046 , B01J2219/00585 , B01J2219/00596 , B01J2219/00722 , C12N15/10 , C12N15/1031 , C12N15/66 , C12P19/34 , C12Q2521/343 , C12Q2521/50 , C12Q2521/501 , 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.
摘要翻译：方法和装置涉及在载体上具有预定序列的多核苷酸的合成。组装方法包括产生重叠构建寡核苷酸的引物延伸和目标多核苷酸组装到锚定载体结合的寡核苷酸上。公开了用于选择具有预定义的序列和/或长度的多核苷酸的方法和装置。

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