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1. 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。

2. 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。

3. 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。

4. 发明申请

US20110311980A1 Nucleic Acid Amplification and Sequencing on a Droplet Actuator 审中-公开
标题翻译：滴液致动器中的核酸扩增和测序
公开(公告)号：US20110311980A1
公开(公告)日：2011-12-22
申请号：US13139467
申请日：2009-12-15
申请人： Michael G. Pollack , Allen E. Eckhardt , Vijay Srinivasan , Prasanna Thwar
发明人： Michael G. Pollack , Allen E. Eckhardt , Vijay Srinivasan , Prasanna Thwar
IPC分类号： C12Q1/68 , C12M1/34
CPC分类号： B01L3/502761 , B01L3/502784 , B01L3/502792 , B01L7/525 , B01L2200/10 , B01L2200/143 , B01L2300/0816 , B01L2300/0838 , B01L2400/0427 , B01L2400/043 , B01L2400/0644 , C12Q1/6869 , C12Q2565/301 , C12Q2565/626
摘要： The invention provides a droplet actuator device, as well as systems, methods and devices making use of the droplet actuator device. The droplet actuator device may include a substrate having electrodes arranged for conducting one or more droplet operations. The droplet actuator device may include a substrate having a reactor path with a wash region associated with a magnet for immobilizing mobilizing beads during bead washing operations. The droplet actuator device may include nucleotide base reservoirs and dedicated nucleotide base electrode paths arranged for transporting nucleotide base droplets from nucleotide base reservoirs to the reactor path. The droplet actuator device may include one or more wash buffer reservoirs associated with electrode paths arranged for transporting wash buffer droplets from wash buffer reservoirs to the reactor path. The droplet actuator device may include one or more sample reservoirs and sample paths arranged for transporting sample droplets from the one or more sample reservoirs to the reactor path. The droplet actuator device may include one or more enzyme reservoirs and dedicated enzyme electrode paths arranged for transporting enzyme droplets from the one or more enzyme reservoirs to a detection electrode.
摘要翻译：本发明提供一种液滴致动器装置，以及利用液滴致动器装置的系统，方法和装置。液滴致动器装置可以包括具有布置用于进行一个或多个液滴操作的电极的基板。液滴致动器装置可以包括具有反应器路径的基底，其具有与磁珠相关联的洗涤区域，用于在珠洗涤操作期间固定动员珠粒。液滴致动器装置可以包括核苷酸碱基储存器和专用核苷酸碱基电极路径，其设置用于将核苷酸碱基液滴从核苷酸碱基储存器输送到反应器路径。液滴致动器装置可以包括一个或多个洗涤缓冲液储存器，其与设置用于将洗涤缓冲液滴从洗涤缓冲液储存器运送到反应器路径的电极路径相关联。液滴致动器装置可以包括一个或多个样品储存器和样品路径，其被布置用于将样品液滴从一个或多个样品储存器输送到反应器路径。液滴致动器装置可以包括一个或多个酶储器和用于将酶液滴从一个或多个酶储存器输送到检测电极的专用酶电极路径。

5. 发明申请

US20100221713A1 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：US20100221713A1
公开(公告)日：2010-09-02
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。

6. 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。

7. 发明申请

US20130225452A1 Method of Preparing a Nucleic Acid Library 审中-公开
公开(公告)号：US20130225452A1
公开(公告)日：2013-08-29
申请号：US13596965
申请日：2012-08-28
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： C12N15/10
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.

8. 发明申请

US20130225450A1 Method of Ligating a Nucleic Acid 审中-公开
标题翻译：核酸的方法
公开(公告)号：US20130225450A1
公开(公告)日：2013-08-29
申请号：US13596548
申请日：2012-08-28
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.
摘要翻译：制备与油接触的液滴核酸文库的方法，包括：（a）在油中的液滴中的平端核酸片段以产生平端核酸片段; （b）使油中的液滴中的平端核酸片段磷酸化以产生磷酸化的核酸片段; 将A尾连接到油中的液滴中的磷酸化核酸片段以产生A尾核酸片段; 和（d）将核酸衔接子偶联到油中的液滴中的A尾核酸片段，以产生包含衔接子连接的核酸片段的核酸文库。

9. 发明授权

US08852952B2 Method of loading a droplet actuator 有权
标题翻译：装载液滴致动器的方法
公开(公告)号：US08852952B2
公开(公告)日：2014-10-07
申请号：US12990815
申请日：2009-05-04
申请人： Michael G. Pollack , Prasanna Thwar , Vamsee K. Pamula , Allen Eckhardt , Alexander Shenderov , Dwayne Allen , Vijay Srinivasan
发明人： Michael G. Pollack , Prasanna Thwar , Vamsee K. Pamula , Allen Eckhardt , Alexander Shenderov , Dwayne Allen , Vijay Srinivasan
IPC分类号： G01N1/00
CPC分类号： B01L3/502784 , B01L3/502715 , B01L3/502761 , B01L7/52 , B01L2200/027 , B01L2200/0673 , B01L2300/022 , B01L2300/023 , B01L2300/024 , B01L2300/0672 , B01L2300/0816 , B01L2300/0864 , B01L2300/0867 , B01L2400/0415 , B01L2400/0424 , B01L2400/0427 , B01L2400/043 , B01L2400/0478 , B01L2400/0481 , B01L2400/0487 , B01L2400/049 , B01L2400/0683 , G01N27/44704 , G01N35/0098 , G01N2035/1046 , Y10T436/25 , Y10T436/2575
摘要： The invention provides droplet actuators and droplet actuator cassettes including reagent storage capabilities, as well as methods of making and using the droplet actuators and cassettes. The invention also provides continuous flow channel elements and techniques for using electrodes to manipulate droplets in flowing streams. The invention also discloses methods of separating compounds on a droplet actuator. Various other aspects of the invention are also disclosed.
摘要翻译：本发明提供了包括试剂储存能力的液滴致动器和液滴致动器盒，以及制造和使用液滴致动器和盒的方法。本发明还提供了用于使用电极在流动流中操纵液滴的连续流动通道元件和技术。本发明还公开了在液滴致动器上分离化合物的方法。还公开了本发明的各种其它方面。

10. 发明授权

US09675972B2 Method of concentrating beads in a droplet 有权
公开(公告)号：US09675972B2
公开(公告)日：2017-06-13
申请号：US12921256
申请日：2009-03-09
申请人： Michael G. Pollack , Ramakrishna Sista , Zhishan Hua , Vamsee K. Pamula , Arjun Sudarsan , Vijay Srinivasan , Prasanna Thwar , Alexander Shenderov , Dwayne Allen
发明人： Michael G. Pollack , Ramakrishna Sista , Zhishan Hua , Vamsee K. Pamula , Arjun Sudarsan , Vijay Srinivasan , Prasanna Thwar , Alexander Shenderov , Dwayne Allen
IPC分类号： B01L3/00 , B01L3/02 , B01F13/00 , B01F13/08 , B03C1/01 , B03C1/28
CPC分类号： B01L3/0282 , B01F13/002 , B01F13/0023 , B01F13/0071 , B01F13/0076 , B01F13/0818 , B01L3/502753 , B01L2200/0621 , B01L2200/0631 , B01L2200/0652 , B01L2200/0668 , B01L2200/141 , B01L2300/0832 , B01L2300/0864 , B01L2300/0867 , B01L2400/043 , B03C1/01 , B03C1/284 , B03C1/286 , B03C2201/18 , B03C2201/26 , B03C2201/28 , Y10T436/25
摘要： Methods of concentrating beads in a droplet and/or loading beads on a fluidic device are provided, including among other things, a method of concentrating beads in a droplet, the method comprising: (a) providing a droplet actuator comprising: (i) an interior droplet operations volume; and (ii) a reservoir exterior to the interior volume; (iii) a droplet established in a liquid path extending from the reservoir into the interior volume; (b) providing magnetically responsive beads in the portion of the droplet which is in the reservoir; (c) magnetically attracting the magnetically responsive beads through the liquid path into the portion of the droplet which is in the interior volume; and (d) forming a droplet comprising one or more of the magnetically responsive beads in the interior volume.

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