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48 条记录 (耗时 0.043 秒)

    Enzymatic Assays for a Droplet Actuator
    31.
    发明申请
    Enzymatic Assays for a Droplet Actuator 有权
    滴液致动器的酶检测

    公开(公告)号：US20100151439A1

    公开(公告)日：2010-06-17

    申请号：US12531844

    申请日：2008-03-23

    申请人： Vamsee K. Pamula ,  Allen E. Eckhardt

    发明人： Vamsee K. Pamula ,  Allen E. Eckhardt

    IPC分类号： C12Q1/00 ,  C12N9/00

    CPC分类号： G01N21/6486 ,  B01F13/0809 ,  C07H15/04 ,  C12Q1/34 ,  G01N2333/928 ,  G01N2800/042

    摘要： A method of conducting a droplet-based enzymatic assay is provided. The method generally makes use of a droplet actuator. A droplet comprising an enzyme of interest is provided on the droplet actuator along with a droplet comprising a substrate which is potentially modified in the presence of the enzyme. The method involves executing droplet operations on the droplet actuator to combine the droplets, thereby yielding an assay droplet, and detecting modification of the substrate by the enzyme in the assay droplet on the droplet actuator. The enzyme of interest may, for example, be a potentially mutated or improperly folded enzyme exhibiting altered enzyme activity as compared to a corresponding normal enzyme.

    摘要翻译： 提供了一种进行基于液滴的酶测定的方法。 该方法通常使用液滴致动器。 包含感兴趣的酶的液滴与包含底物的液滴一起提供在液滴致动器上，所述液滴在酶的存在下可能被修饰。 该方法涉及对液滴致动器执行液滴操作以组合液滴，从而产生测定液滴，并且通过液滴致动器上的测定液滴中的酶检测底物的修饰。 感兴趣的酶可以是例如与相应的正常酶相比显示改变的酶活性的潜在突变或不正确折叠的酶。

    Enzymatic Assays Using Umbelliferone Substrates with Cyclodextrins in Droplets of Oil
    32.
    发明申请
    Enzymatic Assays Using Umbelliferone Substrates with Cyclodextrins in Droplets of Oil 有权
    使用伞形酮底物与环糊精在油滴中的酶检测

    公开(公告)号：US20110118132A1

    公开(公告)日：2011-05-19

    申请号：US13012831

    申请日：2011-01-25

    申请人： Theodore Winger ,  Allen E. Eckhardt ,  Carrie Graham ,  Ramakrishna Sista ,  Vamsee K. Pamula ,  Tong Wang ,  Yalin Xiong ,  Ning Wu ,  Sanjay Saha ,  Gajendrasinh Balvantsinh Raolji ,  Raveendra Dayam

    发明人： Theodore Winger ,  Allen E. Eckhardt ,  Carrie Graham ,  Ramakrishna Sista ,  Vamsee K. Pamula ,  Tong Wang ,  Yalin Xiong ,  Ning Wu ,  Sanjay Saha ,  Gajendrasinh Balvantsinh Raolji ,  Raveendra Dayam

    IPC分类号： C40B30/00 ,  C12Q1/00

    CPC分类号： C12Q1/00

    摘要： The invention is directed to droplet actuator devices and assay methods. The method may include, among other things, a method of conducting enzymatic assays using umbelliferone substrates with cyclodextrins in droplets in oil, the method including incubating a droplet in oil, the droplet preferably comprising an umbelliferone substrate, a sample, and a cyclodextrin compound. The methods may further include a method of substantially eliminating cross-contamination between droplets in enzymatic containing substrate-based bioassays. The method may include immobilization of the enzymatic substrate including forming an inclusion complex with the substrate for stabilizing the substrate within an aqueous environment, wherein the inclusion complex may be formed using cyclodextrins. In yet another embodiment, the invention may provide a method of enhancing hydrolysis of enzymatic substrates. The methods may further include a method of forming an inclusion complex with the substrate for stabilizing the substrate within an aqueous environment, wherein the substrate may be a 4-MU- or HMU containing substrate. The inclusion complex may be formed using cyclodextrin.

    摘要翻译： 本发明涉及液滴致动器装置和测定方法。 该方法尤其可以包括使用具有环糊精的伞形酮底物在油中的液滴进行酶测定的方法，所述方法包括将液滴在油中孵育，所述液滴优选包含伞形酮底物，样品和环糊精化合物。 所述方法还可以包括基本上消除含酶底物生物测定中的液滴之间的交叉污染的方法。 该方法可以包括固定酶底物，包括与底物形成包合物以使其在水性环境中稳定，其中包合物可以使用环糊精形成。 在另一个实施方案中，本发明可提供增强酶底物水解的方法。 所述方法还可以包括在水性环境中与基材形成包含配合物以稳定基材的方法，其中所述基材可以是含有4-MU-或HMU的底物。 包合物可以使用环糊精形成。

    Method of Ligating a Nucleic Acid
    35.
    发明申请
    Method of Ligating a Nucleic Acid 审中-公开
    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。