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1. 发明授权

US08716015B2 Manipulation of cells on a droplet actuator 有权
标题翻译：操纵液滴致动器上的细胞
公开(公告)号：US08716015B2
公开(公告)日：2014-05-06
申请号：US12334575
申请日：2008-12-15
申请人： Michael G. Pollack , Vamsee K. Pamula , Allen E. Eckhardt
发明人： Michael G. Pollack , Vamsee K. Pamula , Allen E. Eckhardt
IPC分类号： C12N5/00 , C12P19/34 , C12Q1/68 , C12Q1/00 , G01N1/10 , G01N1/00 , G01N35/00
CPC分类号： C12N13/00 , B01J2219/00527 , B01J2219/00653 , B01J2219/00743 , G01N15/10 , G01N35/10 , G01N2015/0088 , G01N2015/1081 , G01N2035/00564 , G01N2035/1034 , Y10T436/11 , Y10T436/25 , Y10T436/2575
摘要： A method of inoculating a culture medium including providing a droplet including a single cell type on a droplet actuator and inoculating a culture medium with the droplet. A method of providing a metabolically useful substance to a cell culture, including providing a droplet actuator including a cell culture droplet loaded thereon, the sample droplet including cells and a cell culture medium, and a second droplet comprising a metabolically useful substance. The method also includes conducting one or more droplet operations to combine the cell culture droplet with the second droplet on the droplet actuator. Related methods, droplet actuators, and systems are also provided.
摘要翻译：一种接种培养基的方法，包括在液滴致动器上提供包含单细胞类型的液滴并用液滴接种培养基。提供代谢有用的物质给细胞培养物的方法，包括提供包括其上装载有细胞培养物液滴的液滴致动器，所述样品液滴包括细胞和细胞培养基，以及包含代谢有用物质的第二液滴。该方法还包括进行一个或多个液滴操作以将细胞培养液滴与液滴致动器上的第二液滴组合。还提供了相关方法，液滴致动器和系统。

2. 发明申请

US20130288254A1 Droplet Actuator and Droplet-Based Techniques 审中-公开
标题翻译：滴液致动器和基于液滴的技术
公开(公告)号：US20130288254A1
公开(公告)日：2013-10-31
申请号：US13390121
申请日：2010-08-13
申请人： Michael G. Pollack , Vijay Srinivasan , Allen E. Eckhardt , Prasanna Thwar , Zhishan Hua , Arjun Sudarsan , Theodore Winger
发明人： Michael G. Pollack , Vijay Srinivasan , Allen E. Eckhardt , Prasanna Thwar , Zhishan Hua , Arjun Sudarsan , Theodore Winger
IPC分类号： C12Q1/68
CPC分类号： G01N27/447 , B01L3/502761 , B01L3/502792 , B01L7/52 , B01L2200/0668 , B01L2300/0819 , B01L2300/0864 , B01L2300/0867 , B01L2300/089 , B01L2400/0427 , B01L2400/043 , C12Q1/686 , G01N33/54326 , G01N2035/00425 , G01N2035/1058 , G01N2035/1062 , C12Q2549/101 , C12Q2527/15 , C12Q2527/107 , C12Q2561/113
摘要： The invention is directed to certain droplet actuated molecular techniques. In one embodiment, the invention provides droplet actuator methods for detection of single nucleotide polymorphisms (SNPs) in a DNA sequence using digital microfluidics, including droplet actuator-based sample preparation and SNP analysis. In another embodiment, the invention provides droplet actuator devices and methods for providing integrated sample preparation and multiplexed detection of an infectious agent, such as HIV. In yet another embodiment, the invention provides droplet actuator devices and techniques for PCR amplification and detection of specific nucleic acid sequences using digital microfluidics, including droplet actuator-based sample preparation and target nucleic acid analysis. In yet another embodiment the invention provides methods for performing hot-start PCR on a droplet actuator. In yet another embodiment, the method of the invention combines PCR amplification with pyrosequencing to investigate specific sequences.
摘要翻译：本发明涉及某些液滴致动分子技术。在一个实施方案中，本发明提供用于使用数字微流体（包括基于液滴致动器的样品制备和SNP分析）检测DNA序列中的单核苷酸多态性（SNP）的液滴致动器方法。在另一个实施方案中，本发明提供了液滴致动器装置和用于提供综合样品制备和多重检测感染因子如HIV的方法。在另一个实施方案中，本发明提供了使用数字微流体（包括基于液滴致动器的样品制备和靶核酸分析）的PCR扩增和检测特异性核酸序列的液滴致动器装置和技术。在另一个实施方案中，本发明提供了在液滴致动器上进行热启动PCR的方法。在另一个实施方案中，本发明的方法将PCR扩增与焦磷酸测序结合以研究特异性序列。

3. 发明申请

US20130178374A1 Systems for and Methods of Hybrid Pyrosequencing 有权
公开(公告)号：US20130178374A1
公开(公告)日：2013-07-11
申请号：US13542721
申请日：2012-07-06
申请人： Allen E. Eckhardt , Jonathan Benton , Deborah Boles
发明人： Allen E. Eckhardt , Jonathan Benton , Deborah Boles
IPC分类号： C12N15/10 , C12Q1/68
CPC分类号： C12N15/1089 , C12Q1/6869 , C12Q1/689 , C12Q1/701 , C12Q1/703 , G06F19/22 , G06F19/24 , C12Q2537/165 , C12Q2565/301
摘要： The systems and methods of the invention provide a guided approach to pyrosequencing (i.e., hybrid pyrosequencing). A de novo nucleic acid sequence may compared to a library of possible results and the next nucleotide to be dispensed is selected based on the comparison of the de novo sequence and the library of possible results. In another example, at least the first nucleotide to be dispensed is selected based on a query of a database(s) of non-sequence parameters (e.g., incidence of infection, diagnostic symptoms, sample source) and subsequent dispensations determined based on a comparison of the de novo sequence and the library of possible results (e.g., candidate sequences). The systems and methods of the invention may be performed using a droplet actuator.

4. 发明申请

US20110256542A9 Droplet Actuator Devices, Systems, and Methods 有权
标题翻译：液滴致动器装置，系统和方法
公开(公告)号：US20110256542A9
公开(公告)日：2011-10-20
申请号：US12546115
申请日：2009-08-24
申请人： Michael G. Pollack , Zhishan Hua , Allen E. Eckhardt , Prasanna Thwar , Vijay Srinivasan , Vamsee K. Pamula
发明人： Michael G. Pollack , Zhishan Hua , Allen E. Eckhardt , Prasanna Thwar , Vijay Srinivasan , Vamsee K. Pamula
IPC分类号： C12Q1/68
CPC分类号： B01L3/502792 , B01L3/502753 , B01L3/502761 , B01L7/525 , B01L2200/10 , B01L2300/0816 , B01L2300/089 , B01L2400/0427 , B01L2400/043 , C12Q1/6848 , C12Q2565/629
摘要： The invention relates to certain novel approaches to reducing or eliminating the movement of contaminants from one droplet to another on a droplet actuator via liquid filler fluid. In one application, droplet actuators are used to conduct genetic analysis using polymerase chain reaction (PCR) techniques. The invention addresses the need for improved methods of performing PCR on a droplet actuator that provide for optimum amplification and detection of a sample target.
摘要翻译：本发明涉及在液滴致动器上通过液体填充液减少或消除污染物从一个液滴到另一个液滴的移动的某些新颖方法。在一个应用中，液滴致动器用于使用聚合酶链反应（PCR）技术进行遗传分析。本发明解决了对提供样品靶的最佳放大和检测的在液滴致动器上进行PCR的改进方法的需要。

5. 发明申请

US20100140093A1 Droplet-Based Surface Modification and Washing 有权
标题翻译：基于液滴的表面改性和洗涤
公开(公告)号：US20100140093A1
公开(公告)日：2010-06-10
申请号：US12615609
申请日：2009-11-10
申请人： Vamsee K. Pamula , Vijay Srinivasan , Allen E. Eckhardt , Michael G. Pollack , Richard B. Fair
发明人： Vamsee K. Pamula , Vijay Srinivasan , Allen E. Eckhardt , Michael G. Pollack , Richard B. Fair
IPC分类号： C25B7/00
CPC分类号： G01N27/26 , B01F13/0071 , B01F13/0076 , B01L3/0268 , B01L3/502715 , B01L3/502723 , B01L3/502761 , B01L3/502784 , B01L3/502792 , B01L7/52 , B01L7/525 , B01L2200/027 , B01L2200/0668 , B01L2200/0673 , B01L2200/0684 , B01L2300/0654 , B01L2300/0816 , B01L2300/0861 , B01L2300/0864 , B01L2300/0867 , B01L2300/089 , B01L2300/1827 , B01L2400/0406 , B01L2400/0415 , B01L2400/0424 , B01L2400/0427 , B01L2400/043 , B01L2400/0436 , B01L2400/0442 , B01L2400/0448 , C12Q1/686 , C12Q1/6869 , G01N27/3271 , G01N27/416 , G01N27/447 , G01N35/00029 , G01N2035/00158 , G01N2035/00237
摘要： The present invention relates to droplet-based surface modification and washing. According to one embodiment, a method of providing a bead-containing droplet with a reduced concentration of a substance is provided, the method including providing a droplet microactuator including a droplet having a starting volume and including one or more beads and a starting concentration and starting quantity of the substance. The method further includes conducting one or more droplet operations to merge a wash droplet with the droplet provided in the above step to yield a combined droplet and including one or more droplet operations to divide the combined droplet to yield a set of droplets including a droplet including substantially all of the one or more beads and having a decreased concentration of the substance relative to the starting concentration and a droplet which is substantially lacking in the beads.
摘要翻译：本发明涉及液滴表面改性和洗涤。根据一个实施例，提供了一种提供具有降低的物质浓度的含珠粒的液滴的方法，所述方法包括提供包括具有起始体积并包括一个或多个珠粒和起始浓度的液滴的液滴微致动器，并起始浓度物质的数量。该方法还包括进行一个或多个液滴操作以将洗涤液滴与上述步骤中提供的液滴合并以产生组合液滴并且包括一个或多个液滴操作以分开组合的液滴以产生包括液滴的一组液滴，包括液滴，包括基本上所有的一种或多种珠粒，并且物质相对于起始浓度降低，而珠子中基本上不含有液滴。

6. 发明申请

US20100048410A1 Bead Sorting on a Droplet Actuator 审中-公开
标题翻译：珠子在滴液致动器上排序
公开(公告)号：US20100048410A1
公开(公告)日：2010-02-25
申请号：US12531826
申请日：2008-03-24
申请人： Alexander D. Shenderov , Vamsee K. Pamula , Allen E. Eckhardt
发明人： Alexander D. Shenderov , Vamsee K. Pamula , Allen E. Eckhardt
IPC分类号： C40B30/00 , C40B50/08
CPC分类号： G01N33/54366 , G01N33/54326 , G01N35/0098
摘要： A method of sorting beads on a droplet actuator. The method may, for example, include the following steps: (a) providing a droplet actuator comprising a substrate comprising electrodes arranged for conducting droplet operations on a substrate surface; (b) providing an assay droplet on the substrate surface, the droplet comprising two or more target-capture bead populations comprising target-capture beads comprising: (i) a capture probe bound to a target substance; and (ii) a unique bar binding element which binds to a corresponding binder; (c) using droplet operations to combine the assay droplet with a bead-capture droplet comprising one or more bead-capture beads having affinity for the binding element; (d) immobilizing the one or more bead-capture beads while conducting droplet operations to separate the bead-capture beads from unbound target-capture beads; (e) resuspending the one or more bead-capture beads in a droplet, thereby providing a droplet comprising a substantially pure substance-capture bead population; and (f) using droplet operations to conduct one or more protocol steps for an assay protocol.
摘要翻译：一种在液滴致动器上分选珠粒的方法。该方法可以例如包括以下步骤：（a）提供液滴致动器，其包括基板，该基板包括布置成用于在基板表面上进行液滴操作的电极; （b）在基底表面上提供测定液滴，所述液滴包含两个或更多个目标捕获珠群，包含目标捕获珠，其包含：（i）与靶物质结合的捕获探针; 和（ii）结合相应的粘合剂的独特的棒结合元件; （c）使用液滴操作将测定液滴与包含对结合元件具有亲和性的一个或多个珠粒捕获珠粒的珠粒捕获液滴组合; （d）固定所述一个或多个珠粒捕获珠粒，同时进行液滴操作以将珠粒捕获珠粒与未结合的靶捕获珠粒分离; （e）将一个或多个珠粒捕获珠重新悬浮在液滴中，由此提供包含基本上纯的物质捕获珠粒的液滴; 和（f）使用液滴操作为测定方案进行一个或多个方案步骤。

7. 发明申请

US20090311713A1 Method of Detecting an Analyte 有权
标题翻译：检测分析物的方法
公开(公告)号：US20090311713A1
公开(公告)日：2009-12-17
申请号：US12546253
申请日：2009-08-24
申请人： Michael G. Pollack , Zhishan Hua , Allen E. Eckhardt , Prasanna Thwar , Vijay Srinivasan , Vamsee K. Pamula
发明人： Michael G. Pollack , Zhishan Hua , Allen E. Eckhardt , Prasanna Thwar , Vijay Srinivasan , Vamsee K. Pamula
IPC分类号： C12Q1/68
CPC分类号： B01L3/502792 , B01L3/502753 , B01L3/502761 , B01L7/525 , B01L2200/10 , B01L2300/0816 , B01L2300/089 , B01L2400/0427 , B01L2400/043 , C12Q1/6848 , C12Q2565/629
摘要： The invention relates to certain novel approaches to reducing or eliminating the movement of contaminants from one droplet to another on a droplet actuator via liquid filler fluid. In one application, a method of detecting an analyte is provided and includes providing in a detection window a droplet including a signal-producing substance indicative of the presence and/or quantity of an analyte and one or more magnetically responsive beads which may interfere with signal produced by the signal producing substance. The method further includes using a magnetic field for magnetically removing the magnetically responsive beads from the detection window and/or magnetically restricting the magnetically responsive beads from entering the detection window while transporting and/or retaining the droplet in the detection window. The method additionally includes detecting a signal produced by the signal-producing substance.
摘要翻译：本发明涉及在液滴致动器上通过液体填充液减少或消除污染物从一个液滴到另一个液滴的移动的某些新颖方法。在一个应用中，提供检测分析物的方法，并且包括在检测窗口中提供包含指示分析物的存在和/或量的信号产生物质的液滴和一个或多个可能干扰信号的磁响应珠子由信号产生物质产生。该方法还包括使用磁场来从检测窗口磁性地去除磁响应珠子和/或磁力限制磁响应珠子进入检测窗口，同时在检测窗口中传送和/或保持液滴。该方法还包括检测由信号产生物质产生的信号。

8. 发明申请

US20090069371A1 ISOQUINOLINE COMPOUNDS 审中-公开
标题翻译：异喹啉化合物
公开(公告)号：US20090069371A1
公开(公告)日：2009-03-12
申请号：US12274887
申请日：2008-11-20
申请人： Mitchell A. deLong , Marcos L. Sznaidman , Robert H. Oakley , Allen E. Eckhardt , Christine Hudson , Jeffrey D. Yingling , Michael Peel , Thomas E. Richardson , Clare Louise Murray , Byappanahally N. Narasinga Rao , Brian H. Heasley , Paresma R. Patel
发明人： Mitchell A. deLong , Marcos L. Sznaidman , Robert H. Oakley , Allen E. Eckhardt , Christine Hudson , Jeffrey D. Yingling , Michael Peel , Thomas E. Richardson , Clare Louise Murray , Byappanahally N. Narasinga Rao , Brian H. Heasley , Paresma R. Patel
IPC分类号： A61K31/47 , C07D217/22 , A61P27/02 , A61P9/00 , A61P1/00 , A61P15/08 , A61P35/00 , A61P19/08 , C12N5/00
CPC分类号： C07D217/02
摘要： Isoquinoline compounds with G are provided that influence, inhibit or reduce the action of a G-protein receptor kinase. Pharmaceutical compositions including therapeutically effective amounts of the isoquinoline compounds and pharmaceutically acceptable carriers are also provided. Various methods using the compounds and/or compositions to affect disease states or conditions such as cancer, osteoporosis and glaucoma are also provided.
摘要翻译：提供了具有G的异喹啉化合物，其影响，抑制或降低G蛋白受体激酶的作用。还提供包含治疗有效量的异喹啉化合物和药学上可接受的载体的药物组合物。还提供了使用化合物和/或组合物影响疾病状态或病症如癌症，骨质疏松症和青光眼的各种方法。

9. 发明申请

US20130203606A1 Method of Preparing a Nucleic Acid Library 审中-公开
公开(公告)号：US20130203606A1
公开(公告)日：2013-08-08
申请号：US13581354
申请日：2011-02-22
申请人： Michael G Pollack , Allen E. Eckhardt , Prasanna Thwar , Jeremy Rouse
发明人： Michael G Pollack , Allen E. Eckhardt , Prasanna Thwar , Jeremy Rouse
IPC分类号： B01J19/00
CPC分类号： C12N15/1075 , B01J19/0046 , C12N15/1006 , C40B40/08 , C40B50/06
摘要： A method of preparing a nucleic acid library in droplets in contact with oil, including: (a) blunt-ending nucleic acid fragments in a droplet in the oil to yield blunt-ended nucleic acid fragments; (b) phosphorylating the blunt-ended nucleic acid fragments in a droplet in the oil to yield phosphorylated nucleic acid fragments; coupling A-tails to the phosphorylated nucleic acid fragments in a droplet in the oil to yield A-tailed nucleic acid fragments; and (d) coupling nucleic acid adapters to the A-tailed nucleic acid fragments in a droplet in the oil to yield the nucleic acid library comprising adapter-ligated nucleic acid fragments.

10. 发明申请

US20110097763A1 Thermal Cycling Method 审中-公开
标题翻译：热循环法
公开(公告)号：US20110097763A1
公开(公告)日：2011-04-28
申请号：US12991966
申请日：2009-05-13
申请人： Michael G. Pollack , Zhishan Hua , Allen E. Eckhardt , Prasanna Thwar , Vijay Srinivasan , Vamsee K. Pamula
发明人： Michael G. Pollack , Zhishan Hua , Allen E. Eckhardt , Prasanna Thwar , Vijay Srinivasan , Vamsee K. Pamula
IPC分类号： C12P19/34
CPC分类号： B01L7/525 , B01L3/502761 , B01L3/502792 , B01L7/52 , B01L2200/027 , B01L2200/0668 , B01L2200/10 , B01L2300/0803 , B01L2300/0816 , B01L2300/0861 , B01L2300/0864 , B01L2300/0867 , B01L2300/088 , B01L2300/0883 , B01L2300/089 , B01L2300/1827 , B01L2400/0427 , B01L2400/043 , C12Q1/686 , C12Q2565/629
摘要： The invention relates to certain novel approaches to reducing or eliminating the movement of contaminants from one droplet to another on a droplet actuator via liquid filler fluid. In one application, droplet actuators are used to conduct genetic analysis using polymerase chain reaction (PCR) techniques. The invention addresses the need for improved methods of performing PCR on a droplet actuator that provide for optimum amplification and detection of a sample target.
摘要翻译：本发明涉及在液滴致动器上通过液体填充液减少或消除污染物从一个液滴到另一个液滴的移动的某些新颖方法。在一个应用中，液滴致动器用于使用聚合酶链反应（PCR）技术进行遗传分析。本发明解决了对提供样品靶的最佳放大和检测的在液滴致动器上进行PCR的改进方法的需要。

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