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1. 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。

2. 发明申请

WO2007117256A1 MULTIPLEXED AMPLIFICATION OF SHORT NUCLEIC ACIDS 审中-公开
标题翻译：短期核酸的多重放大
公开(公告)号：WO2007117256A1
公开(公告)日：2007-10-18
申请号：PCT/US2006/021172
申请日：2006-05-31
申请人： APPLERA CORPORATION , LAO, Kai Qin , LIVAK, Kenneth, J. , STRAUS, Neil, A.
发明人： LAO, Kai Qin , LIVAK, Kenneth, J. , STRAUS, Neil, A.
IPC分类号： C12Q1/68 , C12P19/34
CPC分类号： C12N15/1065 , C12Q1/6851 , C12Q1/686 , C12Q2525/301 , C12Q2525/161 , C12Q2563/179 , C12Q2537/143 , C12Q2525/207
摘要： The present teachings provide methods, compositions, and kits for reverse transcribing and amplifying small nucleic acids such as micro RNAs. High levels of multiplexing are provided by the use of a zip-coded stem-loop reverse transcription primer, along with a PCR- based pre-amplification reaction that comprises a zip-coded forward primer. Detector probes in downstream decoding PCRs can take advantage of the zip-code introduced by the stem-loop reverse transcription primer. In some embodiments, further amplification is achieved by cycling the reverse transcription reaction. The present teachings also provide compositions and kits useful for performing the reverse transcription and amplification reactions described herein.
摘要翻译：本教导提供用于逆转录和扩增小核酸如微RNA的方法，组合物和试剂盒。通过使用zip编码的茎环逆转录引物以及包含经翻译的正向引物的基于PCR的预扩增反应提供了高水平的复用。下游解码PCR中的检测器探针可以利用茎环逆转录引物引入的zip码。在一些实施方案中，通过循环逆转录反应来实现进一步的扩增。本教导还提供可用于进行本文所述的逆转录和扩增反应的组合物和试剂盒。

3. 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.

4. 发明申请

WO2007044066A2 METHODS FOR NORMALIZING AND FOR IDENTIFYING SMALL NUCLEIC ACIDS 审中-公开
标题翻译：用于正常化和鉴定小核酸的方法
公开(公告)号：WO2007044066A2
公开(公告)日：2007-04-19
申请号：PCT/US2006/012025
申请日：2006-03-29
申请人： APPLERA CORPORATION , LAO, Kai Qin , STRAUS, Neil, A. , BURNS, John, W.
发明人： LAO, Kai Qin , STRAUS, Neil, A. , BURNS, John, W.
IPC分类号： C12Q1/68 , C12P19/34
CPC分类号： C12Q1/6855 , C12Q1/6851 , C12Q2535/138 , C12Q2525/121 , C12Q2525/301
摘要： The present teachings are generally directed to methods for normalizing at least one species of small nucleic acid that is present in a population of small nucleic acid species, wherein the relative concentration of at least one small nucleic acid species is substantially greater than the relative concentration of at least one other small nucleic acid species in the population. At least one small nucleic acid species is normalized using a multiplicity of primers comprising degenerate sequences. In some embodiments, a small nucleic acid species is identified by inserting at least part of an extension product from a normalized population into a vector and subsequently sequencing the insert. In some embodiments, a small nucleic acid species is identified by determining the sequence of at least part of an extension product.
摘要翻译：本教导通常涉及用于归一化存在于小核酸物种群中的至少一种小核酸的方法，其中至少一种小核酸物质的相对浓度基本上大于群体中至少一种其他小核酸物种。使用包含简并序列的多个引物将至少一个小核酸物质归一化。在一些实施方案中，通过将来自标准化群体的至少部分延伸产物插入载体并随后对插入物进行测序来鉴定小核酸物种。在一些实施方案中，通过确定延伸产物的至少一部分的序列来鉴定小核酸物种。

5. 发明申请

WO2006115570A2 SMALL NUCLEIC ACID DETECTION PROBES AND USES THEREOF 审中-公开
标题翻译：小核酸检测探针及其用途
公开(公告)号：WO2006115570A2
公开(公告)日：2006-11-02
申请号：PCT/US2006/005828
申请日：2006-02-17
申请人： APPLERA CORPORATION , LAO, Kai Qin , STRAUS, Neil A.
发明人： LAO, Kai Qin , STRAUS, Neil A.
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6818 , C12Q2563/173 , C12Q2545/114 , C12Q2525/301
摘要： The present teachings are directed to compositions, methods, and kits for detecting and quantitating small nucleic acid molecules, including small DNA molecules and small RNA molecules. The detector probes of the current teachings, including single-loop detector probes, double-loop detector probes, and bimolecular detector probes, are designed to selectively hybridize with a corresponding small nucleic acid target and to produce, under appropriate conditions, a detectable signal or a detectably different signal. The detector complexes of the current teachings comprise a detector probe comprising a first reporter group and a displaceable sequence comprising a second reporter group, wherein the displaceable sequence is hybridized to the detector probe. According to certain methods, detecting a small nucleic acid target comprises the target displacing the displaceable sequence of a detector complex to form a detector probe-small nucleic acid target duplex, illuminating the duplex with light of an appropriate wavelength, and determining the presence of a detectable fluorescent signal or the change in a detectable signal.
摘要翻译：本教导涉及用于检测和定量小核酸分子（包括小DNA分子和小RNA分子）的组合物，方法和试剂盒。目前的教导的检测器探针，包括单环检测器探针，双环检测器探针和双分子检测器探针被设计为与相应的小核酸靶选择性杂交，并在适当的条件下产生可检测的信号或一个可检测的不同信号。本教导的检测器复合物包括检测器探针，其包括第一报告基团和包含第二报告基团的可置换序列，其中可移位序列与检测器探针杂交。根据某些方法，检测小核酸靶包括靶取代检测器复合物的可置换序列以形成检测器探针 - 小核酸靶双链体，用合适波长的光照亮双链体，并确定可检测的荧光信号或可检测信号的变化。

6. 发明申请

WO2007032817A2 PURE MIRNA SAMPLE PREPARATION METHOD 审中-公开
标题翻译：纯MIRNA样品制备方法
公开(公告)号：WO2007032817A2
公开(公告)日：2007-03-22
申请号：PCT/US2006/027756
申请日：2006-07-17
申请人： APPLERA CORPORATION , LAO, Kai Qin
发明人： LAO, Kai Qin
IPC分类号： C12Q1/68 , C07H21/02 , C12P19/34 , C12N1/08
CPC分类号： C12N15/1003 , C12N1/06 , C12N1/08 , C12N15/111 , C12N2310/14 , C12N2310/315 , C12N2310/321 , C12N2310/341 , C12N2310/346 , C12N2330/10 , C12N2330/30 , C12P19/34 , C12Q1/68 , C12N2310/3521
摘要： The present teachings provide novel methods, compositions, and kits for analyzing mature micro RNAs (miRNAs). By taking advantage of the observation that most mature miRNAs in cells are tightly associated with RISCs, the present teachings provide approaches for studying mature miRNAs without the complications of additional nucleic acids. For example, in some embodiments the present teachings provide a method of purifying mature miRNAs comprising heating a sample to form a lysate, and, degrading the additional nucleic acids. The resulting mixture lacks the additional nucleic acids, and contains mature miRNAs associated with RISCs. Liberating the mature miRNAs from RISCs, for example by a protease, a detergent, and/or heat, can result in a pure collection of mature miRNAs.
摘要翻译：本教导提供用于分析成熟微RNA（miRNA）的新型方法，组合物和试剂盒。通过利用细胞中大多数成熟miRNA与RISC紧密相关的观察，本教导提供了研究成熟miRNA而没有附加核酸并发症的方法。例如，在一些实施方案中，本发明提供纯化成熟miRNA的方法，其包括加热样品以形成裂解物，以及降解另外的核酸。所得混合物缺乏额外的核酸，并且含有与RISC相关的成熟miRNA。从RISC解离成熟的miRNA，例如通过蛋白酶，洗涤剂和/或加热，可导致成熟miRNA的纯收集。

7. 发明申请

WO2005094532A2 ENCODING AND DECODING REACTIONS FOR DETERMINING TARGET POLYNUCLEOTIDES 审中-公开
标题翻译：用于确定目标多核苷酸的编码和解码反应
公开(公告)号：WO2005094532A2
公开(公告)日：2005-10-13
申请号：PCT/US2005/009882
申请日：2005-03-24
申请人： APPLERA CORPORATION , LAO, Kai Qin , REED, Mark T.
发明人： LAO, Kai Qin , REED, Mark T.
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6869 , C12Q1/6851 , C12Q2525/191 , C12Q2525/185 , C12Q2521/501 , C12Q2525/155 , C12Q2521/313 , C12Q2537/143 , C12Q2525/161
摘要： The present invention is directed to methods, reagents, and kits for detecting the presence or absence of (or quantifying) target polynucleotide sequences in at least one sample using encoding and decoding reactions. When a particular target polynucleotide is present in a sample, a reaction product is formed in the encoding reaction that includes addressable primer portions. At least one label and at least one address primer is employed in the decoding amplification reaction thereby providing a detectable signal value depending upon whether a sequence is present or absent. In some embodiments universal bases and reduced libraries of probes can be employed for highly multiplexed analysis of target polynucleotides.
摘要翻译：本发明涉及用于使用编码和解码反应检测至少一个样品中靶多核苷酸序列的存在或不存在（或定量）的方法，试剂和试剂盒。当特定靶多核苷酸存在于样品中时，在编码反应中形成包含可寻址引物部分的反应产物。在解码扩增反应中使用至少一个标签和至少一个地址引物，从而根据序列是否存在提供可检测的信号值。在一些实施方案中，探针的通用碱基和还原文库可用于靶多核苷酸的高度多重分析。

8. 发明申请

WO2012083189A2 METHODS, COMPOSITIONS, SYSTEMS, APPARATUSES AND KITS FOR NUCLEIC ACID AMPLIFICATION 审中-公开
标题翻译：用于核酸扩增的方法，组合物，系统，装置和试剂盒
公开(公告)号：WO2012083189A2
公开(公告)日：2012-06-21
申请号：PCT/US2011/065535
申请日：2011-12-16
申请人： LIFE TECHNOLOGIES CORPORATION , LI, Bin , LAO, Kai Qin , O'NEIL, Jennifer , KUNKEL, Jennifer , HALEY, Kellie , KASINSKAS, Rachel , MA, Zhaochun , BRZOSKA, Pius
发明人： LI, Bin , LAO, Kai Qin , O'NEIL, Jennifer , KUNKEL, Jennifer , HALEY, Kellie , KASINSKAS, Rachel , MA, Zhaochun , BRZOSKA, Pius
IPC分类号： C12Q1/68
CPC分类号： C12Q1/46 , C12P19/34 , C12Q1/6834 , C12Q1/6846 , C12Q1/6853 , C12Q1/6869 , C12Q1/6874 , C12Q2531/119 , C12Q2565/537
摘要： Novel methods of generating a localized population of immobilized clonal amplicons on a support are provided.
摘要翻译：提供了在支持物上产生固定化克隆扩增子的局部群体的新方法。

9. 发明公开

EP1917367A2 METHODS FOR RNA PROFILING 审中-公开
标题翻译：方法RNA配置文件创建
公开(公告)号：EP1917367A2
公开(公告)日：2008-05-07
申请号：EP06800092.6
申请日：2006-07-17
申请人： Applera Corporation
发明人： LIVAK, Kenneth , LAO, Kai Qin
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6883 , C12Q1/6809 , C12Q1/6876 , C12Q2600/158 , C12Q2600/178 , C12Q2525/207
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.

10. 发明公开

EP1831400A2 ID-TAG COMPLEXES, ARRAYS, AND METHODS OF USE THEREOF 审中-公开
标题翻译： ID标签复杂，数组和及其使用方法
公开(公告)号：EP1831400A2
公开(公告)日：2007-09-12
申请号：EP05853238.3
申请日：2005-12-06
申请人： Applera Corporation
发明人： SCHROTH, Gary P. , LAO, Kai Qin , STRAUS, Neil A.
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6823 , C12Q1/6816 , C12Q2565/519 , C12Q2537/1373 , C12Q2537/125 , C12Q2565/501 , C12Q2563/131 , C12Q2525/197 , C12Q2563/185
摘要： The present invention relates to the detection of target sequences. Detection can be achieved through the use of ID-tag complexes. These ID-tag complexes are relatively stable in the absence of a target sequence. In the presence of a target sequence, the complexes dissociate and form new complexes or duplexes, which can be purified or eliminated and detected on an ID-tag system.

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