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    Methods for normalizing and for identifying small nucleic acids
    2.
    发明申请
    Methods for normalizing and for identifying small nucleic acids 审中-公开
    归一化和鉴定小核酸的方法

    公开(公告)号：US20060223066A1

    公开(公告)日：2006-10-05

    申请号：US11093587

    申请日：2005-03-29

    申请人： Kai Lao ,  Neil Straus ,  John Burns

    发明人： Kai Lao ,  Neil Straus ,  John Burns

    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。

    Ligation assay
    4.
    发明申请
    Ligation assay 有权
    连接试验

    公开(公告)号：US20050064459A1

    公开(公告)日：2005-03-24

    申请号：US10866012

    申请日：2004-06-10

    申请人： Kai Lao

    发明人： Kai Lao

    IPC分类号： C12Q1/68

    CPC分类号： C12Q1/682 ,  C12Q1/6827 ,  C12Q2561/125 ,  C12Q2531/113 ,  C12Q2525/155 ,  C12Q2525/125

    摘要： Disclosed are methods and kits for detecting or quantifying one or more target polynucleotide sequences in a sample. Embodiments of the invention employ a first ligation reaction, a subsequent optional amplification reaction, and a second ligation reaction. Embodiments of the invention combine the specificity of both hybridization and ligation reactions along with universal probe polynucleotide sequences to achieve specific and multiplexed detection of a plurality of target polynucleotide sequences.

    摘要翻译： 公开了用于检测或定量样品中一种或多种靶多核苷酸序列的方法和试剂盒。 本发明的实施方案采用第一连接反应，随后任选的扩增反应和第二连接反应。 本发明的实施方案将杂交和连接反应的特异性与通用探针多核苷酸序列结合，以实现多个靶多核苷酸序列的特异性和多重检测。

    MULTIPLEXED AMPLIFICATION OF SHORT NUCLEIC ACIDS
    5.
    发明申请
    MULTIPLEXED AMPLIFICATION OF SHORT NUCLEIC ACIDS 审中-公开
    短期核酸的多重放大

    公开(公告)号：US20070077570A1

    公开(公告)日：2007-04-05

    申请号：US11421449

    申请日：2006-05-31

    申请人： Kai Lao ,  Kenneth Livak ,  Neil Straus

    发明人： Kai Lao ,  Kenneth Livak ,  Neil Straus

    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。

    Pure miRNA Sample Preparation Method
    6.
    发明申请
    Pure miRNA Sample Preparation Method 审中-公开
    纯miRNA样品制备方法

    公开(公告)号：US20070059739A1

    公开(公告)日：2007-03-15

    申请号：US11458077

    申请日：2006-07-17

    申请人： Kai Lao

    发明人： Kai Lao

    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，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。

    Compositions, methods, and kits for identifying and quantitating small RNA molecules
    7.
    发明申请
    Compositions, methods, and kits for identifying and quantitating small RNA molecules 审中-公开
    用于鉴定和定量小RNA分子的组合物，方法和试剂盒

    公开(公告)号：US20060057595A1

    公开(公告)日：2006-03-16

    申请号：US10944153

    申请日：2004-09-16

    申请人： Kai Lao ,  Neil Straus

    发明人： Kai Lao ,  Neil Straus

    IPC分类号： C12Q1/68 ,  C07H21/02 ,  C12P19/34

    CPC分类号： C12Q1/6851 ,  C12Q2525/207 ,  C12Q2525/161 ,  C12Q2521/107

    摘要： Compositions, methods, and kits for identifying and for quantitating polynucleotide targets are disclosed. These compositions, methods, and kits are particularly useful when the polynucleotide target is a small RNA molecule, including without limitation microRNA (miRNA), small interfering RNA (siRNA), and certain other classes of non-coding RNA molecules. The forward and reverse primers of the disclosed first primer sets comprise unusually short target-binding portions that are 6-10 nucleotides long. Certain of the disclosed methods employ one or more multiplex reaction steps to identify, quantitate, or identify and quantitate, a multiplicity of target polynucleotides.

    摘要翻译： 公开了用于鉴定和定量多核苷酸靶标的组合物，方法和试剂盒。 当多核苷酸靶是小RNA分子时，这些组合物，方法和试剂盒特别有用，包​​括但不限于microRNA（miRNA），小干扰RNA（siRNA）和某些其他类型的非编码RNA分子。 所公开的第一引物组的正向和反向引物包含6-10个核苷酸长的异常短的靶结合部分。 某些所公开的方法采用一个或多个多重反应步骤来鉴定，定量或鉴定和定量多个靶多核苷酸。

    Analog probe complexes
    9.
    发明申请
    Analog probe complexes 审中-公开
    模拟探针复合物

    公开(公告)号：US20060014191A1

    公开(公告)日：2006-01-19

    申请号：US11172280

    申请日：2005-06-29

    申请人： Kai Lao ,  Timothy Geiser ,  Neil Straus

    发明人： Kai Lao ,  Timothy Geiser ,  Neil Straus

    IPC分类号： C12Q1/68 ,  C07K14/47

    CPC分类号： B82Y10/00 ,  B82Y5/00 ,  C12Q1/6832 ,  C12Q1/6876 ,  C12Q2525/101

    摘要： The present invention relates to the detection of target sequences. The present description discloses compositions and methods involving analog nucleic acids, such as PNA and L-DNA, for the detection of nucleic acids. Additionally, hybrid detectable markers are provided.

    摘要翻译： 本发明涉及靶序列的检测。 本说明书公开了涉及用于检测核酸的类似物核酸如PNA和L-DNA的组合物和方法。 另外，提供了杂交可检测标记。