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1. 发明授权

US08206953B2 Message abundance and allele copy number determination using IVT with single-stranded primer-promoter-selector constructs 有权
标题翻译：使用IVT与单链引物启动子选择构建体的消息丰度和等位基因拷贝数确定
公开(公告)号：US08206953B2
公开(公告)日：2012-06-26
申请号：US11525064
申请日：2006-09-21
申请人： Nataliya Korzheva , Michael Seul
发明人： Nataliya Korzheva , Michael Seul
IPC分类号： C12P19/34
CPC分类号： C12Q1/6837 , C12N15/1096 , C12Q1/6858 , C12Q1/6865 , C12Q1/6876 , C12Q2521/107 , C12Q2525/143 , C12Q2563/179 , C12Q2565/137 , C12Q2545/114 , C12Q2531/113
摘要： Disclosed is a single stranded primer-promoter-selector construct comprising (in 3′ to 5′ orientation) a primer subsequence annealing to the target, a T7 or other promoter subsequence (the template strand), and a selector subsequence. The primer can be extended by template mediated elongation, including reverse transcription, or ligation to another oligonucleotide. The promoter sequence is oriented to direct the in-vitro transcription (IVT) opposite to that of primer extension, where the selector subsequence serves as a template for IVT. The selector is associated with the target subsequence of interest and it, and the amplified product are unique subsequences, dissimilar to other sequence present in the sample. The construct's is useful for determination of the presence and relative abundance of designated subsequences in the sample, multiplex gene expression analysis, multiplex allele counting, determination of polymorphic/mutation site, and loss of heterozygosity.
摘要翻译：公开了一种单链引物 - 启动子选择构建体，其包含（靶向3'至5'取向）引物亚序列退火，T7或其它启动子亚序列（模板链）和选择子序列。引物可通过模板介导的延伸扩增，包括逆转录或连接至另一寡核苷酸。启动子序列被定向以引导与引物延伸相反的体外转录（IVT），其中选择子序列用作IVT的模板。选择器与感兴趣的目标子序列相关联，并且扩增产物是与样本中存在的其他序列不同的唯一子序列。该构建体可用于确定样品中指定的子序列的存在和相对丰度，多重基因表达分析，多重等位基因计数，多态/突变位点的确定和杂合性的丧失。

2. 发明申请

US20060188896A1 Nucleic acid amplification with integrated multiplex dectection 有权
标题翻译：核酸扩增与综合多重检测
公开(公告)号：US20060188896A1
公开(公告)日：2006-08-24
申请号：US11218838
申请日：2005-09-02
申请人： Michael Seul , Nataliya Korzheva , Jiacheng Yang , Yi Zhang
发明人： Michael Seul , Nataliya Korzheva , Jiacheng Yang , Yi Zhang
IPC分类号： C12Q1/68 , C12P19/34
CPC分类号： C12Q1/6865 , C12Q2563/149 , C12Q2563/107 , C12Q2537/143
摘要： A method mediated with in-vitro transcription (“IVT”) which permits miniaturization of multiplexed DNA and RNA analysis, and in which elongation-mediated multiplexed analysis of polymorphisms (eMAP®) is used as the analysis step, is described. Also described is a method mediated with IVT is for selecting a designated strand from T7-tagged double stranded DNA: wherein, the selected strand forms the template for RNA synthesis. In one embodiment, double stranded DNA incorporating the T7 (or other) promoter sequence at the 3′ end or the 5′ end is produced, for example, by amplification of genomic DNA using the Polymerase Chain Reaction (PCR). Also disclosed are nested PCR designs permitting allele analysis in combination with strand selection by IVT. Further, in one embodiment of a homogeneous format for transcription-mediated amplification and multiplexed detection (which may be particularly suited for viral or pathogen detection), encoded microparticles display “looped” capture probe configurations permitting the generation of a signal upon capture of RNA product and real-time assay monitoring.
摘要翻译：描述了允许多元化DNA和RNA分析的小型化以及其中使用延长介导的多态性多重分析（eMAP））介导的体外转录（“IVT”）介导的方法作为分析步骤。还描述了用IVT介导的用于从T7标记的双链DNA中选择指定链的方法：其中所选择的链形成用于RNA合成的模板。在一个实施方案中，例如通过使用聚合酶链式反应（PCR）扩增基因组DNA来产生在3'末端或5'端掺入T7（或其它）启动子序列的双链DNA。还公开了允许等位基因分析与IVT的链选择组合的巢式PCR设计。此外，在用于转录介导的扩增和多重检测（其可能特别适用于病毒或病原体检测）的均匀形式的一个实施方案中，编码的微粒显示允许在捕获RNA产物时产生信号的“环状”捕获探针构型和实时检测监测。

3. 发明申请

US20120094298A1 NUCLEIC ACID AMPLIFICATION WITH INTEGRATED MULTIPLEX DETECTION 审中-公开
标题翻译：具有集成多重检测的核酸扩增
公开(公告)号：US20120094298A1
公开(公告)日：2012-04-19
申请号：US13179136
申请日：2011-07-08
申请人： Michael Seul , Nataliya Korzheva , Jiacheng Yang , Yi Zhang
发明人： Michael Seul , Nataliya Korzheva , Jiacheng Yang , Yi Zhang
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6865
摘要： A method mediated with in-vitro transcription (“IVT”) which permits miniaturization of multiplexed DNA and RNA analysis, and in which elongation-mediated multiplexed analysis of polymorphisms (eMAP®) is used as the analysis step, is described. Also described is a method mediated with IVT is for selecting a designated strand from T7-tagged double stranded DNA: wherein, the selected strand forms the template for RNA synthesis. In one embodiment, double stranded DNA incorporating the T7 (or other) promoter sequence at the 3′ end or the 5′end is produced, for example, by amplification of genomic DNA using the Polymerase Chain Reaction (PCR). Also disclosed are nested PCR designs permitting allele analysis in combination with strand selection by IVT. Further, in one embodiment of a homogeneous format for transcription-mediated amplification and multiplexed detection (which may be particularly suited for viral or pathogen detection), encoded microparticles display “looped” capture probe configurations permitting the generation of a signal upon capture of RNA product and real-time assay monitoring.
摘要翻译：描述了使用允许多重DNA和RNA分析的小型化的体外转录（“IVT”）介导的方法，并且其中使用延长介导的多态性复合分析（eMAP）作为分析步骤。还描述了用IVT介导的用于从T7标记的双链DNA中选择指定链的方法：其中所选择的链形成用于RNA合成的模板。在一个实施方案中，例如通过使用聚合酶链式反应（PCR）扩增基因组DNA来产生在3'端或5'端掺入T7（或其它）启动子序列的双链DNA。还公开了允许等位基因分析与IVT的链选择组合的巢式PCR设计。此外，在用于转录介导的扩增和多重检测（其可能特别适用于病毒或病原体检测）的均匀形式的一个实施方案中，编码的微粒显示允许在捕获RNA产物时产生信号的“环状”捕获探针构型和实时检测监测。

4. 发明授权

US07977050B2 Nucleic acid amplification with integrated multiplex detection 有权
标题翻译：核酸扩增与综合多重检测
公开(公告)号：US07977050B2
公开(公告)日：2011-07-12
申请号：US11218838
申请日：2005-09-02
申请人： Michael Seul , Nataliya Korzheva , Jiacheng Yang , Yi Zhang
发明人： Michael Seul , Nataliya Korzheva , Jiacheng Yang , Yi Zhang
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6865 , C12Q2563/149 , C12Q2563/107 , C12Q2537/143
摘要： A method mediated with in-vitro transcription (“IVT”) which permits miniaturization of multiplexed DNA and RNA analysis, and in which elongation-mediated multiplexed analysis of polymorphisms (eMAP®) is used as the analysis step, is described. Also described is a method mediated with IVT is for selecting a designated strand from T7-tagged double stranded DNA: wherein, the selected strand forms the template for RNA synthesis. In one embodiment, double stranded DNA incorporating the T7 (or other) promoter sequence at the 3′ end or the 5′ end is produced, for example, by amplification of genomic DNA using the Polymerase Chain Reaction (PCR). Also disclosed are nested PCR designs permitting allele analysis in combination with strand selection by IVT. Further, in one embodiment of a homogeneous format for transcription-mediated amplification and multiplexed detection (which may be particularly suited for viral or pathogen detection), encoded microparticles display “looped” capture probe configurations permitting the generation of a signal upon capture of RNA product and real-time assay monitoring.
摘要翻译：描述了使用允许多重DNA和RNA分析的小型化的体外转录（“IVT”）介导的方法，并且其中使用延长介导的多态性复合分析（eMAP）作为分析步骤。还描述了用IVT介导的用于从T7标记的双链DNA中选择指定链的方法：其中所选择的链形成用于RNA合成的模板。在一个实施方案中，例如通过使用聚合酶链式反应（PCR）扩增基因组DNA来产生在3'末端或5'端掺入T7（或其它）启动子序列的双链DNA。还公开了允许等位基因分析与IVT的链选择组合的巢式PCR设计。此外，在用于转录介导的扩增和多重检测（其可能特别适用于病毒或病原体检测）的均匀形式的一个实施方案中，编码的微粒显示允许在捕获RNA产物时产生信号的“环状”捕获探针构型和实时检测监测。

5. 发明申请

US20130040840A1 NUCLEIC ACID AMPLIFICATION WITH INTEGRATED MULTIPLEX DETECTION 审中-公开
标题翻译：具有集成多重检测的核酸扩增
公开(公告)号：US20130040840A1
公开(公告)日：2013-02-14
申请号：US13540211
申请日：2012-07-02
申请人： Hui Huang , Sukanta BANERJEE , Michael Seul , Nataliya Korzheva , Jiacheug Yang , Yi Zhang
发明人： Hui Huang , Sukanta BANERJEE , Michael Seul , Nataliya Korzheva , Jiacheug Yang , Yi Zhang
IPC分类号： C40B30/04 , C40B60/12 , C40B40/06
CPC分类号： C12Q1/6804 , G01N33/6803 , G01N33/6854 , G01N2333/705 , G01N2458/10 , C12Q2563/179
摘要： Compositions and methods of detecting multiple proteins of interest in a sample using arrays are provided herein.
摘要翻译：本文提供了使用阵列检测样品中感兴趣的多种蛋白质的组合物和方法。

6. 发明申请

US20070065864A1 Message abundance and allele copy number determination using IVT with single-stranded primer-promoter-selector constructs 有权
标题翻译：使用IVT与单链引物启动子选择构建体的消息丰度和等位基因拷贝数确定
公开(公告)号：US20070065864A1
公开(公告)日：2007-03-22
申请号：US11525064
申请日：2006-09-21
申请人： Nataliya Korzheva , Michael Seul
发明人： Nataliya Korzheva , Michael Seul
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6837 , C12N15/1096 , C12Q1/6858 , C12Q1/6865 , C12Q1/6876 , C12Q2521/107 , C12Q2525/143 , C12Q2563/179 , C12Q2565/137 , C12Q2545/114 , C12Q2531/113
摘要： Disclosed is a single stranded primer-promoter-selector construct comprising (in 3′ to 5′ orientation) a primer subsequence annealing to the target, a T7 or other promoter subsequence (the template strand), and a selector subsequence. The primer can be extended by template mediated elongation, including reverse transcription, or ligation to another oligonucleotide. The promoter sequence is oriented to direct the in-vitro transcription (IVT) opposite to that of primer extension, where the selector subsequence serves as a template for IVT. The selector is associated with the target subsequence of interest and it, and the amplified product are unique subsequences, dissimilar to other sequence present in the sample. The construct's is useful for determination of the presence and relative abundance of designated subsequences in the sample, multiplex gene expression analysis, multiplex allele counting, determination of polymorphic/mutation site, and loss of heterozygosity.
摘要翻译：公开了一种单链引物 - 启动子选择构建体，其包含（靶向3'至5'取向）引物亚序列退火，T7或其它启动子亚序列（模板链）和选择子序列。引物可通过模板介导的延伸扩增，包括逆转录或连接至另一寡核苷酸。启动子序列被定向以引导与引物延伸相反的体外转录（IVT），其中选择子序列用作IVT的模板。选择器与感兴趣的目标子序列相关联，并且扩增产物是与样本中存在的其他序列不同的唯一子序列。该构建体可用于确定样品中指定的子序列的存在和相对丰度，多重基因表达分析，多重等位基因计数，多态/突变位点的确定和杂合性的丧失。

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