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1. 发明申请

US20110251083A1 Multiplexed Amplification of Short Nucleic Acids 审中-公开
标题翻译：短核酸的多重扩增
公开(公告)号：US20110251083A1
公开(公告)日：2011-10-13
申请号：US12762272
申请日：2010-04-16
申请人： Kai Qin LAO , Kenneth J. Livak , Neil A. Straus
发明人： Kai Qin LAO , Kenneth J. Livak , Neil A. Straus
IPC分类号： C40B30/04 , C12P19/34 , C40B40/06 , C12Q1/68
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的预扩增反应来提供高水平的复用。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。

2. 发明申请

US20100317062A1 HOT START REVERSE TRANSCRIPTION BY PRIMER DESIGN 有权
标题翻译：通过PRIMER设计开始反向转录
公开(公告)号：US20100317062A1
公开(公告)日：2010-12-16
申请号：US12504633
申请日：2009-07-16
申请人： Kai Qin LAO , Neil A. Straus , Kenneth J. Livak
发明人： Kai Qin LAO , Neil A. Straus , Kenneth J. Livak
IPC分类号： C12P19/34 , C12N9/12 , C07H21/00
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端自补偿干杆的热起始引物，并且循环和延伸产物在高温下形成，但是在低温下减少延伸产物形成。

3. 发明申请

US20130184171A1 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：US20130184171A1
公开(公告)日：2013-07-18
申请号：US13615057
申请日：2012-09-13
申请人： Kai Qin LAO , Kenneth J. Livak , Neil A. Straus
发明人： Kai Qin LAO , Kenneth J. Livak , Neil A. Straus
IPC分类号： C12N15/10
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的预扩增反应来提供高水平的复用。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。

4. 发明申请

US20100221790A1 Analyzing Messenger RNA and Micro RNA in the Same Reaction Mixture 有权
标题翻译：在相同的反应混合物中分析Messenger RNA和微RNA
公开(公告)号：US20100221790A1
公开(公告)日：2010-09-02
申请号：US12781690
申请日：2010-05-17
申请人： Kai Qin LAO , Neil A. Straus
发明人： Kai Qin LAO , Neil A. Straus
IPC分类号： C12P19/34 , C12N9/00
CPC分类号： C12Q1/686 , C12Q1/6809 , C12Q1/6844 , C12Q1/6848 , C12Q1/6853 , C12Q2549/101 , C12Q2537/143 , C12Q2527/107 , C12Q2525/301 , C12Q2525/207 , C12Q2533/101
摘要： The present teachings provide methods, compositions, and kits for performing primer extension reactions on at least two target polynucleotides in the same reaction mixture. In some embodiments, a reverse transcription reaction is performed on a first target polynucleotide with a hot start primer comprising a self-complementary stem and a loop, and extension products form at high temperatures but extension products form less so at low temperatures since the self-complementary stem of the hot start primer prevents hybridization of the target specific region to the target. However, non-hot start primers with free target specific regions can hybridize to their corresponding targets at the low temperature and extension can happen at the low temperature.
摘要翻译：本教导提供用于在相同反应混合物中的至少两个靶多核苷酸上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，对具有包含自身互补的茎和环的热起始底物的第一靶多核苷酸进行逆转录反应，并且在高温下形成延伸产物，但是延伸产物在低温下形成较少，热启动引物的互补干扰物可以防止目标特异性区域与靶标的杂交。然而，具有游离目标特异性区域的非热启动引物可以在低温下与其相应的靶标杂交，并且延伸可以在低温下发生。

5. 发明申请

US20120196292A1 SEQUENCE AMPLIFICATION WITH LINEAR PRIMERS 审中-公开
标题翻译：使用线性预处理器的序列放大
公开(公告)号：US20120196292A1
公开(公告)日：2012-08-02
申请号：US13333419
申请日：2011-12-21
申请人： Kai Qin LAO , Nan Lan Xu , Neil A. Straus
发明人： Kai Qin LAO , Nan Lan Xu , Neil A. Straus
IPC分类号： C12Q1/68 , C12P19/34
CPC分类号： C12Q1/6853 , C12Q1/6844 , C12Q1/6848 , C12Q2600/158 , C12Q2525/301 , C12Q2525/207 , C12Q2525/161
摘要： The present disclosure relates to the amplification of target nucleic acid sequences for various sequencing and/or identification techniques. The use of these primers, as described herein, allows for the reduction in the amplification of nonspecific hybridization events (such as primer dimerization) while allowing for the amplification of the target nucleic acid sequences.
摘要翻译：本公开涉及用于各种测序和/或鉴定技术的靶核酸序列的扩增。如本文所述，这些引物的使用允许减少非特异性杂交事件（例如引物二聚化）的扩增，同时允许靶核酸序列的扩增。

6. 发明申请

US20090171078A1 METHOD OF SEQUENCING NUCLEIC ACIDS USING ELABORATED NUCLEOTIDE PHOSPHOROTIOLATE COMPOUNDS 审中-公开
标题翻译：使用制备的核酸磷酸酯化合物测序核酸的方法
公开(公告)号：US20090171078A1
公开(公告)日：2009-07-02
申请号：US12275176
申请日：2008-11-20
申请人： Kai Qin LAO , Neil A. Straus
发明人： Kai Qin LAO , Neil A. Straus
IPC分类号： C07H19/04 , C12P19/34 , C12Q1/68
CPC分类号： C12Q1/6883
摘要： The present teachings provide methods, compositions, and kits for synthesizing and sequencing nucleic acids. In some embodiments, elaborated nucleotide phosphorothiolate compounds are employed along with efficient cleaving reactions. Improved sequencing efficiency is achieved by the rapid polymerase-mediated incorporation of elaborated nucleotide phosphorothiolate compounds. Increased sequencing efficiency is also achieved by the ability of the cleaving reactions to restore the incorporated nucleotides to their natural structure prior to subsequent elongation.
摘要翻译：本教导提供了用于合成和测序核酸的方法，组合物和试剂盒。在一些实施方案中，精制的核苷酸硫代磷酸酯化合物与有效的切割反应一起使用。通过快速聚合酶介导的拟合的核苷酸硫代硫酸酯化合物的掺入可以改善测序效率。增加的测序效率也可以通过切割反应在随后的伸长之前将掺入的核苷酸恢复到其天然结构的能力来实现。

7. 发明申请

US20100136547A1 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：US20100136547A1
公开(公告)日：2010-06-03
申请号：US12554606
申请日：2009-09-04
申请人： Kai Qin LAO
发明人： Kai Qin LAO
IPC分类号： C12Q1/68 , C12P19/34
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，包括加热样品以形成裂解物，并降解其它核酸。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。

8. 发明申请

US20110159548A1 METHOD FOR AMPLIFYING MONOMORPHIC-TAILED NUCLEIC ACIDS 审中-公开
标题翻译：用于放大单晶尾核的方法
公开(公告)号：US20110159548A1
公开(公告)日：2011-06-30
申请号：US12727202
申请日：2010-03-18
申请人： Kai Qin LAO
发明人： Kai Qin LAO
IPC分类号： C12P19/34
CPC分类号： C12P19/34 , C12Q1/6848 , C12Q2521/107 , C12Q2525/155 , C12Q2525/173
摘要： The present teachings provide methods for amplifying a plurality of target nucleic acids. In some embodiments, a first oligo-dT-universal primer comprising a 3′ oligo-dT portion and a first 5′ universal portion is used to reverse transcribe a plurality of 3′ poly-A tail-containing nucleic acids. A poly-A tail is added to the 3′ end of the first strand products to form a two-tailed reaction product. The two-tailed reaction product is amplified in a PCR, wherein the PCR comprises the first oligo-dT-universal primer, and a second oligo-dT-universal primer, wherein the second oligo-dT-universal primer comprises a 3′ oligo-dT portion and a second 5′ universal portion, wherein the second 5′ universal portion of the second oligo-dT-universal primer comprises a different sequence than the first 5′ universal portion of the first oligo-dT-universal primer. The present teachings also provide compositions and kits for amplifying target nucleic acids containing monomorphic tails.
摘要翻译：本教导提供了扩增多个靶核酸的方法。在一些实施方案中，使用包含3'oligo-dT部分和第一个5'通用部分的第一寡聚-DT通用引物逆转录数多个含3'多聚氨基酸的核酸。将poly-A尾添加到第一链产物的3'末端以形成双尾反应产物。双链反应产物在PCR中扩增，其中PCR包含第一寡聚-D-通用引物和第二寡聚-DT通用引物，其中第二寡聚-D-通用引物包含3'寡聚 - dT部分和第二5'通用部分，其中第二寡聚-D-通用引物的第二个5'通用部分包含与第一寡聚-DT通用引物的前5'通用部分不同的序列。本教导还提供用于扩增含有单形尾的靶核酸的组合物和试剂盒。

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