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1. 发明申请

WO2009015290A1 SYSTEMS AND METHODS FOR ISOLATING NUCLEIC ACIDS 审中-公开
标题翻译：用于分离核酸的系统和方法
公开(公告)号：WO2009015290A1
公开(公告)日：2009-01-29
申请号：PCT/US2008/071071
申请日：2008-07-24
申请人： APPLIED BIOSYSTEMS INC. , VANN, Charles , BREVNOV, Maxim , RUFF, David , LIVAK, Kenneth
发明人： VANN, Charles , BREVNOV, Maxim , RUFF, David , LIVAK, Kenneth
IPC分类号： C12N15/10 , G01N27/447 , B01D57/02
CPC分类号： C12N15/101 , G01N27/44782
摘要： A system for collecting target nucleic acids from a sample can include at least one sample chamber configured to receive a sample containing target nucleic acids and other material, at least one collection chamber removably mountable relative to the at least one sample chamber and configured to collect target nucleic acids separated from the other material, a filter removably mountable relative to the at least one sample chamber and configured to be disposed between the at least one sample chamber and the at least one collection chamber when the at least one collection chamber is mounted relative to the at least one sample chamber. The system may further include a pair of electrodes configured to generate an electric field sufficient to cause target nucleic acids in the at least one sample chamber to migrate via electrophoresis from the at least one sample chamber through the filter into the at least one collection chamber, wherein the filter may be configured to permit passage of target nucleic acids and to block passage of material of a size larger than the target nucleic acids.
摘要翻译：用于从样品收集目标核酸的系统可以包括至少一个样品室，其被配置为接收含有靶核酸和其它材料的样品，至少一个收集室，其相对于至少一个样品室可拆卸地安装并且被配置成收集目标与其他材料分离的核酸，相对于至少一个样品室可拆卸地安装的过滤器，并且被配置为当所述至少一个收集室相对于所述至少一个收集室安装时，被设置在所述至少一个样品室和所述至少一个收集室之间所述至少一个样品室。该系统可以进一步包括一对电极，其被配置为产生足以使所述至少一个样品室中的靶核酸经由电泳从所述至少一个样品室通过所述过滤器迁移到所述至少一个收集室中的电场，其中所述过滤器可以被配置为允许靶核酸的通过并阻止尺寸大于所述靶核酸的材料的通过。

2. 发明申请

WO2007011902A3 METHODS FOR RNA PROFILING 审中-公开
公开(公告)号：WO2007011902A3
公开(公告)日：2007-01-25
申请号：PCT/US2006/027754
申请日：2006-07-17
申请人： APPLERA CORPORATION , LAO, Kai Qin , LIVAK, Kenneth
发明人： LAO, Kai Qin , LIVAK, Kenneth
IPC分类号： C12Q1/68
摘要： The present teachings provide methods, compositions, and kits for detecting micro RNAs (miRNAs). In some embodiments, the miRNAs are quantified from formalin-fixed paraffin-embedded tissue samples in which messenger RNA is degraded. The present teachings take advantage of the observation that most mature miRNAs in vivo are protected by degradation as a result of their association with RISC. Thus, novel methods of studying nucleic acids in archived tissues containing degraded messenger RNA are provided, wherein RISC-protected miRNAs are liberated, and analyzed.

3. 发明申请

WO2007011901A2 HOT START REVERSE TRANSCRIPTION BY PRIMER DESIGN 审中-公开
标题翻译：通过PRIMER设计开始反转转录
公开(公告)号：WO2007011901A2
公开(公告)日：2007-01-25
申请号：PCT/US2006/027753
申请日：2006-07-17
申请人： APPLERA CORPORATION , LAO, Kai Qin , STRAUS, Neil , LIVAK, Kenneth
发明人： LAO, Kai Qin , STRAUS, Neil , LIVAK, Kenneth
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6876 , C07H21/02 , C07H21/04 , C12N9/00 , C12Q1/6844 , C12Q1/6848 , C12Q1/686 , C12Q2549/101 , C12Q2533/101 , C12Q2525/301
摘要： The present teachings provide methods, compositions, and kits for performing primer extension reactions. In some embodiments, a reverse transcription reaction is performed on a target polynucleotide with a hot start primer comprising a blunt-ended self-complementary stem, and a loop, and extension products form at high temperatures but reduce extension product formation at low temperatures.
摘要翻译：本教导提供了用于进行引物延伸反应的方法，组合物和试剂盒。在一些实施方案中，对靶多核苷酸进行逆转录反应，所述靶多核苷酸具有包含平末端自互补茎的热起始引物，并且在高温下形成环，并延伸产物在低温下形成延伸产物。

4. 发明申请

WO2005029041A2 HIGH DENSITY SEQUENCE DETECTION METHODS AND APPARATUS 审中-公开
标题翻译：高密度序列检测方法和装置
公开(公告)号：WO2005029041A2
公开(公告)日：2005-03-31
申请号：PCT/US2004/030805
申请日：2004-09-17
申请人： APPLERA CORPORATION , WOUDENBERG, Timothy , JONES, Bob , LIVAK, Kenneth , LIU, Timothy
发明人： WOUDENBERG, Timothy , JONES, Bob , LIVAK, Kenneth , LIU, Timothy
IPC分类号： G01N
CPC分类号： B01L3/50851 , B01L3/50853 , B01L3/50857 , B01L2200/0642 , B01L2300/044 , B01L2300/0819 , B01L2300/0829 , B01L2400/0409 , B01L2400/0487 , C12Q1/6837
摘要： Methods for amplifying polynucleotides, e.g., by PCR, in a sample comprising polynucleotide targets present at very low concentration, comprising: (a) applying amplification reactants to the surface of a substrate comprising reaction spots, wherein the reactants comprise the sample and an amplification reagent; (b) forming a sealed reaction chamber, having a volume less than about 120 nanoliters, preferably less than about 20 nanoliters, over each of said reaction spots; and (c) thermal cycling the substrate and reactants. In one embodiment, the forming step comprises loading a sealing fluid, e.g., mineral oil, on the surface so as to cover the reaction spots. The present invention also provides microplates, comprising: (a) a substrate having at least about 10,000 reaction spots, each comprising a primer and a droplet of reagent having a volume less than about 120 nanoliters, preferably less then about 20 nanoliters; and (b) a sealing liquid isolating each of the spots.
摘要翻译：在包含以非常低浓度存在的多核苷酸靶标的样品中通过PCR扩增多核苷酸的方法包括：（a）将扩增反应物施加到包含反应点的底物表面，其中反应物包含样品和扩增试剂 ; （b）在每个所述反应点上形成体积小于约120纳升，优选小于约20纳升的密封反应室; 和（c）使底物和反应物热循环。在一个实施例中，成形步骤包括在表面上装载密封流体，例如矿物油，以便覆盖反应点。本发明还提供了微孔板，其包括：（a）具有至少约10,000个反应点的底物，每个底物包含体积小于约120纳升，优选小于约20纳升的底漆和试剂滴; 和（b）隔离每个斑点的密封液体。

5. 发明申请

WO2012054933A2 UNIVERSAL PROBE ASSAY METHODS 审中-公开
标题翻译：通用探针分析方法
公开(公告)号：WO2012054933A2
公开(公告)日：2012-04-26
申请号：PCT/US2011/057536
申请日：2011-10-24
申请人： FLUIDIGM CORPORATION , LIVAK, Kenneth , WEST, Jason A., A. , JONES, Robert, C.
发明人： LIVAK, Kenneth , WEST, Jason A., A. , JONES, Robert, C.
IPC分类号： C12Q1/68
CPC分类号： C12Q1/682 , C12Q1/6876 , C12Q2600/178 , C12Q2525/155 , C12Q2525/307 , C12Q2537/161
摘要： Reagents and methods are provided for detecting the presence of a target polynucleotide in a sample are disclosed. In one aspect, a method for producing a labeled amplification product by amplifying a target nucleic acid sequence to produce an amplification product comprising the target sequence, a first probe-binding sequence 5' to the target sequence, and a second probe-binding sequence 3' to the target sequence, thereby producing an amplification product; and hybridizing a first detection probe to the amplification product, said first detection probe comprising a first segment that hybridizes to the first probe-binding sequence and a second segment that hybridizes to the second probe-binding sequence, thereby producing a labeled amplification product is disclosed.
摘要翻译：提供了用于检测样品中靶多核苷酸存在的试剂和方法。一方面，通过扩增靶核酸序列以产生包含靶序列，与靶序列5'的第一探针结合序列和第二探针结合序列3的扩增产物来产生标记的扩增产物的方法 '到靶序列，从而产生扩增产物; 并将第一检测探针与扩增产物杂交，所述第一检测探针包含与第一探针结合序列杂交的第一片段和与第二探针结合序列杂交的第二片段，从而产生标记的扩增产物

6. 发明申请

WO2008018855A2 METHODS AND KITS FOR EVALUATING DNA METHYLATION 审中-公开
标题翻译：评估DNA甲基化的方法和试剂盒
公开(公告)号：WO2008018855A2
公开(公告)日：2008-02-14
申请号：PCT/US2006/023669
申请日：2006-06-16
申请人： APPLERA CORPORATION , BOYD, Victoria , LIVAK, Kenneth , ZON, Gerald
发明人： BOYD, Victoria , LIVAK, Kenneth , ZON, Gerald
IPC分类号： C12Q1/68 , C12P19/34
CPC分类号： C12Q1/686 , C12Q2565/125 , C12Q2523/125
摘要： Methods and kits are disclosed for determining the degree of methylation of at least one target region. Typically a sample is exposed to a modifying agent to obtain a modified sample comprising a modified nucleotide. At least one target region in the modified sample is amplified. Some of the disclosed methods comprise at least one additional amplification reaction. In some embodiments, at least one mobility shifting analog is incorporated into an amplicon during an amplification reaction. The analogs are analyzed and the degree of methylation of at least one target region is determined.
摘要翻译：公开了用于确定至少一个目标区域的甲基化程度的方法和试剂盒。通常将样品暴露于改性剂以获得包含修饰的核苷酸的修饰样品。修饰样品中的至少一个靶区域被扩增。一些所公开的方法包括至少一个额外的扩增反应。在一些实施方案中，在扩增反应期间将至少一种迁移率移动模拟物掺入扩增子。分析类似物并确定至少一个目标区域的甲基化程度。

7. 发明公开

EP2185705A1 SYSTEMS AND METHODS FOR ISOLATING NUCLEIC ACIDS 审中-公开
标题翻译：系统和方法分离核酸
公开(公告)号：EP2185705A1
公开(公告)日：2010-05-19
申请号：EP08782338.1
申请日：2008-07-24
申请人： Applied Biosystems Inc.
发明人： VANN, Charles , BREVNOV, Maxim , RUFF, David , LIVAK, Kenneth
IPC分类号： C12N15/10 , G01N27/447 , B01D57/02
CPC分类号： C12N15/101 , G01N27/44782
摘要： A system for collecting target nucleic acids from a sample can include at least one sample chamber configured to receive a sample containing target nucleic acids and other material, at least one collection chamber removably mountable relative to the at least one sample chamber and configured to collect target nucleic acids separated from the other material, a filter removably mountable relative to the at least one sample chamber and configured to be disposed between the at least one sample chamber and the at least one collection chamber when the at least one collection chamber is mounted relative to the at least one sample chamber. The system may further include a pair of electrodes configured to generate an electric field sufficient to cause target nucleic acids in the at least one sample chamber to migrate via electrophoresis from the at least one sample chamber through the filter into the at least one collection chamber, wherein the filter may be configured to permit passage of target nucleic acids and to block passage of material of a size larger than the target nucleic acids.

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