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公开(公告)号：US20130225452A1
公开(公告)日：2013-08-29
申请号：US13596965
申请日：2012-08-28
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.
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公开(公告)号：US20130225450A1
公开(公告)日：2013-08-29
申请号：US13596548
申请日：2012-08-28
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。
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公开(公告)号：US20130203606A1
公开(公告)日：2013-08-08
申请号：US13581354
申请日：2011-02-22
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.
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公开(公告)号：US08202686B2
公开(公告)日：2012-06-19
申请号：US12497677
申请日：2009-07-05
IPC分类号： C12Q1/00
CPC分类号： C12Q1/34 , B01F13/0071 , B01L3/502784 , B01L7/54 , B01L2300/0654 , B01L2300/0829 , B01L2300/0867 , B01L2300/089 , B01L2300/1827 , G01N2021/035 , G01N2333/924 , G01N2800/042
摘要： The invention relates to a microfluidic platform and methods of using the platform for conducting enzyme assays using a droplet actuator. The enzyme assays of the invention are useful for, among other things, identifying and/or characterizing disorders resulting from conditions in which enzymes are defective or are produced in inappropriate amounts. Enzyme assays of the invention may, for example, be used to detect altered activity of a particular enzyme in a sample, which may serve as an indicator of a particular disease. Altered activity may, for example, be caused by conditions which result in the increased or reduced production of a certain enzyme or its substrate and/or conditions which result in defective enzymes and/or substrates exhibiting increased or decreased effectiveness relative to corresponding normal enzymes and/or substrates.
摘要翻译： 本发明涉及一种微流体平台和使用该平台用于使用液滴致动器进行酶测定的方法。 本发明的酶测定法尤其可用于识别和/或表征由酶缺陷或以不适当量产生的病症引起的疾病。 本发明的酶测定可以例如用于检测样品中特定酶的改变的活性，其可以用作特定疾病的指标。 改变的活性可以例如由导致某些酶或其底物的产生增加或减少的条件引起，和/或导致有缺陷的酶和/或底物相对于相应的正常酶显示出增加或降低的有效性的条件，和 /或底物。
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公开(公告)号：US08828655B2
公开(公告)日：2014-09-09
申请号：US13862587
申请日：2013-04-15
IPC分类号： C12Q1/00
CPC分类号： G01N21/6486 , B01F13/0809 , C07H15/04 , C12Q1/34 , G01N2333/928 , G01N2800/042
摘要： A method is provided for conducting a droplet-based enzymatic assay, e.g., for diagnostic purposes. On a droplet actuator, a droplet comprising an enzyme of interest is provided along with a droplet comprising a substrate which is potentially modified in the presence of the enzyme. Droplet operations are executed to combine the enzyme and substrate droplets on the droplet actuator, thereby yielding an assay droplet on the droplet actuator. Detecting modification of the substrate by the enzyme in the assay droplet occurs on the droplet actuator. Modified substrate preparations for conducting such enzymatic assays are also provided.
摘要翻译： 提供了用于进行基于液滴的酶测定的方法，例如用于诊断目的。 在液滴致动器上，包含感兴趣的酶的液滴与包含在酶存在下潜在地被修饰的底物的液滴一起提供。 执行液滴操作以将酶和底物液滴组合在液滴致动器上，从而在液滴致动器上产生测定液滴。 在液滴致动器中检测由测定液滴中的酶对底物的修饰。 还提供了用于进行这种酶测定的修饰底物制剂。
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公开(公告)号：US20130230875A1
公开(公告)日：2013-09-05
申请号：US13862587
申请日：2013-04-15
IPC分类号： G01N21/64
CPC分类号： G01N21/6486 , B01F13/0809 , C07H15/04 , C12Q1/34 , G01N2333/928 , G01N2800/042
摘要： A method is provided for conducting a droplet-based enzymatic assay, e.g., for diagnostic purposes. On a droplet actuator, a droplet comprising an enzyme of interest is provided along with a droplet comprising a substrate which is potentially modified in the presence of the enzyme. Droplet operations are executed to combine the enzyme and substrate droplets on the droplet actuator, thereby yielding an assay droplet on the droplet actuator. Detecting modification of the substrate by the enzyme in the assay droplet occurs on the droplet actuator. Modified substrate preparations for conducting such enzymatic assays are also provided.
摘要翻译： 提供了用于进行基于液滴的酶测定的方法，例如用于诊断目的。 在液滴致动器上，包含感兴趣的酶的液滴与包含在酶存在下潜在地被修饰的底物的液滴一起提供。 执行液滴操作以将酶和底物液滴组合在液滴致动器上，从而在液滴致动器上产生测定液滴。 在液滴致动器中检测由测定液滴中的酶对底物的修饰。 还提供了用于进行这种酶测定的修饰底物制剂。
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公开(公告)号：US20100041086A1
公开(公告)日：2010-02-18
申请号：US12497677
申请日：2009-07-05
IPC分类号： C12Q1/34
CPC分类号： C12Q1/34 , B01F13/0071 , B01L3/502784 , B01L7/54 , B01L2300/0654 , B01L2300/0829 , B01L2300/0867 , B01L2300/089 , B01L2300/1827 , G01N2021/035 , G01N2333/924 , G01N2800/042
摘要： The invention relates to a microfluidic platform and methods of using the platform for conducting enzyme assays using a droplet actuator. The enzyme assays of the invention are useful for, among other things, identifying and/or characterizing disorders resulting from conditions in which enzymes are defective or are produced in inappropriate amounts. Enzyme assays of the invention may, for example, be used to detect altered activity of a particular enzyme in a sample, which may serve as an indicator of a particular disease. Altered activity may, for example, be caused by conditions which result in the increased or reduced production of a certain enzyme or its substrate and/or conditions which result in defective enzymes and/or substrates exhibiting increased or decreased effectiveness relative to corresponding normal enzymes and/or substrates.
摘要翻译： 本发明涉及一种微流体平台和使用该平台用于使用液滴致动器进行酶测定的方法。 本发明的酶测定法尤其可用于识别和/或表征由酶缺陷或以不适当量产生的病症引起的疾病。 本发明的酶测定可以例如用于检测样品中特定酶的改变的活性，其可以用作特定疾病的指标。 改变的活性可以例如由导致某些酶或其底物的产生增加或减少的条件引起，和/或导致有缺陷的酶和/或底物相对于相应的正常酶显示出增加或降低的有效性的条件，和 /或底物。
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公开(公告)号：US20110104725A1
公开(公告)日：2011-05-05
申请号：US12990766
申请日：2009-05-04
申请人： Vamsee K. Pamula , Sitaram Emani , Allen Eckhardt , Carrie Graham , Jeremy Rouse
发明人： Vamsee K. Pamula , Sitaram Emani , Allen Eckhardt , Carrie Graham , Jeremy Rouse
IPC分类号： G01N33/53
CPC分类号： G01N33/86 , B01F13/0071 , B01F13/0076 , B01L3/502792 , B01L7/52 , B01L2200/0605 , B01L2200/0668 , B01L2300/0816 , B01L2300/0819 , B01L2300/089 , B01L2400/0406 , B01L2400/0409 , B01L2400/0424 , B01L2400/0427 , B01L2400/043 , B01L2400/046 , B01L2400/0487 , G01N33/5302
摘要： The invention provides techniques for coagulating blood on a droplet actuator. The invention also provides methods of manipulating the coagulated blood including a variety of droplet operations that may be conducted using the coagulated blood. Further, the invention provides a variety of assays that make use of the coagulated blood or various blood samples as input.
摘要翻译： 本发明提供了用于在液滴致动器上凝结血液的技术。 本发明还提供了操作凝血血液的方法，包括可以使用凝血血液进行的各种液滴操作。 此外，本发明提供使用凝血血液或各种血液样品作为输入的各种测定法。
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公开(公告)号：US08440392B2
公开(公告)日：2013-05-14
申请号：US12531844
申请日：2008-03-23
IPC分类号： C12Q1/00
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。
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