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    Droplet actuator devices and methods employing magnetic beads
    6.
    发明授权
    Droplet actuator devices and methods employing magnetic beads 有权

    公开(公告)号：US10379112B2

    公开(公告)日：2019-08-13

    申请号：US14719731

    申请日：2015-05-22

    Applicant: Advanced Liquid Logic, Inc. ,  Duke University

    Inventor： Ramakrishna Sista ,  Vamsee K. Pamula ,  Michael G. Pollack ,  Vijay Srinivasan ,  Allen E. Eckhardt ,  Richard B. Fair

    IPC: B01L3/00 ,  G01N27/447 ,  G01N33/543 ,  C12M1/12 ,  G01N35/10 ,  G01N35/00

    Abstract: A method of providing a droplet in contact with a magnetically responsive bead and having a reduced quantity of a substance. The method generally includes the steps of (a) providing a droplet actuator comprising: (i) a substrate comprising electrodes arranged for conducting droplet operations on a surface; (ii) a starting droplet comprising: (1) one or more magnetically responsive beads; (2) a starting quantity of the substance; and (3) a starting volume; (b) magnetically immobilizing the one or more magnetically responsive beads at a location which is at a distance from a target droplet splitting zone; (c) conducting one or more droplet operations comprising droplet dividing operations selected to divide the combined droplet to yield a set of droplets comprising: (i) a droplet comprising substantially all of the one or more magnetically responsive beads and having a decreased quantity of the substance relative to the starting concentration; and (ii) a droplet substantially lacking the magnetically responsive beads.

    Multiplexed PCR and Fluorescence Detection on a Droplet Actuator
    10.
    发明申请
    Multiplexed PCR and Fluorescence Detection on a Droplet Actuator 审中-公开
    在滴液致动器上进行多路PCR和荧​​光检测

    公开(公告)号：US20140005066A1

    公开(公告)日：2014-01-02

    申请号：US13932719

    申请日：2013-07-01

    Applicant: ADVANCED LIQUID LOGIC INC.

    Inventor： Deborah Boles ,  Lisa Perkins ,  Allen E. Eckhardt ,  Jennifer Foley

    IPC: C12Q1/70 ,  C12Q1/68

    CPC classification number: C12Q1/703 ,  C12Q1/686 ,  C12Q1/701 ,  C12Q2537/143 ,  C12Q2561/113 ,  C12Q2563/107 ,  C12Q2563/159 ,  C12Q2565/102

    Abstract: The present invention provides a droplet actuator device and methods for multiplexed PCR amplification and detection of target amplicons within a single droplet. The methods of the invention combine quantitative real-time PCR (qPCR) amplification with fluorescence-based sequence specific detection technologies for amplified DNA. In one embodiment, fluorescently-labeled oligonucleotide probes may be used for hybridization-based multiplexed detection of target amplicons. The methods of the invention generally involve combining the necessary reactants to form a PCR-ready droplet and thermal cycling the droplet at temperatures sufficient to result in amplification of one or more target nucleic acids. Fluorescence-based detection techniques may be used for end-point or real-time analysis of DNA amplification. For end-point analysis, the accumulation of a signal, e.g., a fluorescence signal, is measured after the amplification of the target sequence is complete. For real-time analysis, the signal is measured while the amplification reaction is in progress.

    Abstract translation: 本发明提供了一种液滴致动器装置和用于在单个液滴内多重PCR扩增和检测靶扩增子的方法。 本发明的方法将定量实时PCR（qPCR）扩增与扩增DNA的基于荧光的序列特异性检测技术相结合。 在一个实施方案中，荧光标记的寡核苷酸探针可用于靶扩增子的杂交多重检测。 本发明的方法通常包括组合必需的反应物以形成PCR准备的液滴并在足以导致一种或多种靶核酸扩增的温度下热循环该液滴。 基于荧光的检测技术可用于DNA扩增的终点或实时分析。 对于终点分析，在靶序列的扩增完成之后测量信号的累积，例如荧光信号。 对于实时分析，在扩增反应进行期间测量信号。

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