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31 条记录 (耗时 0.027 秒)

    Linear amplification of short nucleic acids
    1.
    发明授权
    Linear amplification of short nucleic acids 有权
    短核酸的线性扩增

    公开(公告)号：US07556943B2

    公开(公告)日：2009-07-07

    申请号：US11421319

    申请日：2006-05-31

    申请人： Will Bloch

    发明人： Will Bloch

    IPC分类号： C12P19/34 ,  C12Q1/68

    CPC分类号： C12P19/34 ,  C12Q1/6844 ,  C12Q1/6853 ,  C12Q2531/101 ,  C12Q2525/301 ,  C12Q2521/107 ,  C12Q2527/125

    摘要： The present teachings provide novel methods for amplifying short nucleic acids. In some embodiments, the present teachings provide novel methods for linearly amplifying a collection of micro RNAs by using temperature cycling during a reverse transcription reaction. The cycling can comprise at least 20 cycles of an annealing temperature segment of 10 C-30 C, and a denaturation temperature segment of 35 C-60 C. In some embodiments, the temperature cycled reaction can comprise an osmolyte.

    摘要翻译： 本教导提供了用于扩增短核酸的新方法。 在一些实施方案中，本发明提供了用于在逆转录反应期间通过使用温度循环线性扩增微RNA集合的新方法。 循环可以包括至少20个循环的10℃-30℃的退火温度段和35℃-60℃的变性温度段。在一些实施方案中，温度循环反应可包含渗透剂。

    Detection of mutations in nucleic acids by chemical cleavage
    3.
    发明授权
    Detection of mutations in nucleic acids by chemical cleavage 有权
    通过化学裂解检测核酸中的突变

    公开(公告)号：US5972618A

    公开(公告)日：1999-10-26

    申请号：US130258

    申请日：1998-08-06

    申请人： Will Bloch

    发明人： Will Bloch

    IPC分类号： G01N33/50 ,  C12N15/09 ,  C12Q1/68 ,  C07H21/02 ,  C07H21/04 ,  C12P19/34

    CPC分类号： C12Q1/6827

    摘要： Improved methods, compositions and kits for detecting mutations in nucleic acids are disclosed, such comprise annealing a piece of control nucleic acid without mutations to a piece of test nucleic acid very similar in sequence to the control nucleic acid but possibly containing mutations, treating this mixture with potassium permanganate or hydroxylamine to remove mismatched bases from the duplex nucleic acid, treating the resulting nucleic acid with any of a class of diamines, triamines, and tetraamines analogous to 1,2-ethylenediamine to cleave abasic sites, and then analyzing the chemically treated nucleic acid to determine whether cleavage has occurred and approximately at what position in the nucleic acid any cleavage has occurred. Inclusion of betaine in the base-removal reactions improves their sensitivity and specificity. This series of non-hazardous, non-noxious, chemical treatments can be performed rapidly in a single reaction tube with no or minimal intervening separation steps; and the cleavage product can be prepared for the analysis step without removing the chemical reagents used to effect cleavage.

    摘要翻译： 公开了改进的用于检测核酸突变的方法，组合物和试剂盒，这包括对一片对照核酸进行退火，而不突变到与对照核酸非常相似的测试核酸片段，但可能含有突变，处理该混合物 用高锰酸钾或羟胺从双链核酸中除去错配的碱基，用类似于1,2-乙二胺的一类二胺，三胺和四胺处理所得核酸，以切割脱碱基位点，然后分析化学处理 核酸，以确定是否已经发生切割，并且大约在核酸中发生任何切割的位置。 在脱碱反应中包含甜菜碱可以提高其敏感性和特异性。 这种无毒无毒化学处理可以在单个反应管中快速进行，没有或最小的介入分离步骤; 并且可以制备裂解产物用于分析步骤而不除去用于切割的化学试剂。

    Ionic compounds containing the cationic meriquinone of a benzidine
    6.
    发明授权
    Ionic compounds containing the cationic meriquinone of a benzidine 失效
    含有联苯胺阳离子季戊四醇的离子化合物

    公开(公告)号：US4789630A

    公开(公告)日：1988-12-06

    申请号：US896677

    申请日：1986-08-20

    申请人： Will Bloch ,  Patrick J. Sheridan ,  Robert J. Goodson

    发明人： Will Bloch ,  Patrick J. Sheridan ,  Robert J. Goodson

    IPC分类号： G01N33/545 ,  C12Q1/28 ,  C12Q1/68 ,  G01N21/82 ,  G01N33/535 ,  G01N33/543 ,  G01N33/58 ,  G01N33/53

    CPC分类号： C12Q1/28 ,  C12Q1/68 ,  G01N33/581 ,  C12Q2326/12 ,  Y10S435/803 ,  Y10S435/81 ,  Y10S435/96 ,  Y10S435/975

    摘要： Useful for visualizing biological materials in a solid phase, on a gel, or in a liquid phase is a solid salt of the meriquinone of benzidine or a substituted benzidine. An immobilized or dissolved complex of a polymeric anion and the meriquinone of benzidine or a substituted benzidine having controllable solubility may also be employed. Preferred are meriquinone salts and complexes of 3,3,5,5'-tetramethylbenzidine. For visualization, the benzidine or substituted benzidine is oxidized to its meriquinone at pH 3 to 7 in the presence of an effective anion or polymeric anion, an oxidation catalyst, and an effective amount of oxidant to form a solid salt or immobilized complex of the meriquinone under conditions where the meriquinone solubility lies below about 10.sup.-5 M.

    摘要翻译： 可用于在固相，凝胶或液相中可视化生物材料是联苯胺或取代的联苯胺的固醇的固体盐。 还可以使用固定或溶解的聚合阴离子的复合物和具有可控的溶解度的联苯胺或取代的联苯胺的金刚酮。 优选3,3,5,5'-四甲基联苯胺的季戊四醇盐和络合物。 为了可视化，联苯胺或取代的联苯胺在有效阴离子或聚合阴离子，氧化催化剂和有效量的氧化剂的存在下，在pH 3至7下被氧化成其三聚物，以形成固体盐或固定的复合物 在meriquinone溶解度低于约10 -5 M的条件下。

    Methods for the Reduction of Stutter in Microsatellite Amplification
    7.
    发明申请
    Methods for the Reduction of Stutter in Microsatellite Amplification 有权

    公开(公告)号：US20130011839A1

    公开(公告)日：2013-01-10

    申请号：US13611974

    申请日：2012-09-12

    申请人： Sulekha Rao Coticone ,  Will Bloch

    发明人： Sulekha Rao Coticone ,  Will Bloch

    IPC分类号： C12Q1/68 ,  C12N9/12 ,  C12P19/34

    CPC分类号： C12Q1/686 ,  C12Q1/6846 ,  C12Q1/6848 ,  C12Q1/6876 ,  C12Q2527/125 ,  C12Q2525/151

    摘要： The invention provides a method for reducing stutter in the amplification of a microsatellite comprising the steps of providing a sample comprising a microsatellite having a G+C content of 50% or less; contacting the sample with at least one enzyme having nucleic acid polymerase activity; and incubating the sample with the enzyme for a sufficient amount of time and under conditions sufficient to amplify the microsatellite; wherein the incubation is performed in the presence of an amount of betaine, sorbitol or mixtures thereof, effective to reduce stutter relative to the amount of stutter observed in the absence of betaine and/or sorbitol. The invention also provides compositions containing betaine and/or sorbitol, kits for amplifying microsatellites having a G+C content of 50% or less, and methods of using all of the foregoing.

    Methods for the Reduction of Stutter in Microsatellite Amplification
    8.
    发明申请
    Methods for the Reduction of Stutter in Microsatellite Amplification 审中-公开
    减少微卫星放大技术的方法

    公开(公告)号：US20110143352A1

    公开(公告)日：2011-06-16

    申请号：US12769577

    申请日：2010-04-28

    申请人： Sulekha Rao COTICONE ,  Will Bloch

    发明人： Sulekha Rao COTICONE ,  Will Bloch

    IPC分类号： C12Q1/68 ,  C12N9/12

    CPC分类号： C12Q1/686 ,  C12Q1/6846 ,  C12Q1/6848 ,  C12Q1/6876 ,  C12Q2527/125 ,  C12Q2525/151

    摘要： The invention provides a method for reducing stutter in the amplification of a microsatellite comprising the steps of providing a sample comprising a microsatellite having a G+C content of 50% or less; contacting the sample with at least one enzyme having nucleic acid polymerase activity; and incubating the sample with the enzyme for a sufficient amount of time and under conditions sufficient to amplify the microsatellite; wherein the incubation is performed in the presence of an amount of betaine, sorbitol or mixtures thereof, effective to reduce stutter relative to the amount of stutter observed in the absence of betaine and/or sorbitol. The invention also provides compositions containing betaine and/or sorbitol, kits for amplifying microsatellites having a G+C content of 50% or less, and methods of using all of the foregoing.

    摘要翻译： 本发明提供了减少微卫星扩增中的口吃的方法，包括以下步骤：提供包含G + C含量为50％或更少的微卫星的样品; 使样品与至少一种具有核酸聚合酶活性的酶接触; 并将样品与酶温育足够的时间和足以扩增微卫星的条件; 其中在一定量的甜菜碱，山梨糖醇或其混合物的存在下进行培养，其有效地相对于在不存在甜菜碱和/或山梨醇的情况下观察到的口吃量减少口吃。 本发明还提供含有甜菜碱和/或山梨糖醇的组合物，用于扩增G + C含量为50％以下的微卫星的试剂盒，以及使用上述所有方法。

    METHODS FOR CHARACTERIZING CELLS USING AMPLIFIED MICRO RNAS
    9.
    发明申请
    METHODS FOR CHARACTERIZING CELLS USING AMPLIFIED MICRO RNAS 审中-公开
    使用放大的微RNA RNA表征细胞的方法

    公开(公告)号：US20070048757A1

    公开(公告)日：2007-03-01

    申请号：US11421460

    申请日：2006-05-31

    申请人： Kai Lao ,  Neil Straus ,  Will Bloch

    发明人： Kai Lao ,  Neil Straus ,  Will Bloch

    IPC分类号： C12Q1/68 ,  C12P19/34

    CPC分类号： C12Q1/6809 ,  C12Q1/6853 ,  C12Q2537/143 ,  C12Q2531/113 ,  C12Q2525/207 ,  C12Q2533/101 ,  C12Q2525/301

    摘要： The present teachings provide methods, compositions, and kits for reverse transcribing and amplifying small nucleic acids such as micro RNAs. In some embodiments, the present teachings provide methods of forming micro RNA signatures from single cells, including stem cells. In some embodiments, the present teachings provide methods for determining the identity and/or purity of cells. The present employ performing a multiplexed reverse transcription reaction comprising stem-loop reverse transcription primers, which optionally undergoes temperature cycling, followed by a multiplexed PCR-based pre-amplification reaction, and a subsequently a plurality of lower-plex decoding PCRs.

    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。 在一些实施方案中，本教导提供从单细胞（包括干细胞）形成微RNA特征的方法。 在一些实施方案中，本教导提供了确定细胞的身份和/或纯度的方法。 本发明采用多重逆转录反应，其包括茎环逆转录引物，其可任选进行温度循环，随后进行基于多重PCR的预扩增反应，随后进行多个较低级解码PCR。

    Procaryotic xylose isomerase muteins
    10.
    发明授权
    Procaryotic xylose isomerase muteins 失效
    原核木糖异构酶突变蛋白

    公开(公告)号：US5041378A

    公开(公告)日：1991-08-20

    申请号：US84479

    申请日：1987-08-11

    申请人： Robert J. Drummond ,  Will Bloch ,  Brian W. Matthews ,  Pamela L. Toy

    发明人： Robert J. Drummond ,  Will Bloch ,  Brian W. Matthews ,  Pamela L. Toy

    IPC分类号： C12N9/36 ,  C12N9/92 ,  C12P21/02

    CPC分类号： C12N9/2462 ,  C12N9/92 ,  C12P21/02 ,  Y10S435/886

    摘要： Xylose isomerase (XI) muteins useful in the conversion of glucose to fructose or xylose to xylulose are obtained in usable amounts by protein structural and recombinant DNA methods, including x-ray crystallography, cloning, computer graphic modeling and site-directed mutagenesis and expression of the bacterial DNA sequences encoding native procaryotic xylose isomerase. These native sequences are altered to encode the xylose isomerase muteins having improved catalytic function and/or thermostability.

    摘要翻译： 通过蛋白质结构和重组DNA方法获得可用于将葡萄糖转化为果糖或木糖至木酮糖的木糖异构酶（XI）突变蛋白，包括x射线晶体学，克隆，计算机图形建模和定点诱变和表达 编码天然原核木糖异构酶的细菌DNA序列。 改变这些天然序列以编码具有改善的催化功能和/或热稳定性的木糖异构酶突变蛋白。