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    REAGENTS AND METHODS FOR PCR
    1.
    发明申请
    REAGENTS AND METHODS FOR PCR 有权
    反应物和PCR方法

    公开(公告)号：US20120088275A1

    公开(公告)日：2012-04-12

    申请号：US13256038

    申请日：2010-03-11

    申请人： Lawrence J. Wangh ,  John Rice ,  Nicholas Rice ,  Yanwei Jia

    发明人： Lawrence J. Wangh ,  John Rice ,  Nicholas Rice ,  Yanwei Jia

    IPC分类号： C12P19/34 ,  C12N9/12

    CPC分类号： C12Q1/6806 ,  C12Q1/6853 ,  C12Q2545/114 ,  C12Q2531/107 ,  C12Q2527/107

    摘要： Modified double-stranded oligonucleotides that have terminal regions on each of their strands, that have a hybrid length of 6-50 nucleotides long, that have a melting temperature Tm of at least 32° C., and that include 2-4 modifying groups, each covalently attached to a different terminal region, preferably to a terminal nucleotide, said modifying groups being polycyclic substituents that do not have bulky portions that are non-planar, said modified olgonucleotide being capable of binding to the 5′ ex-nuclease domains of DNA polymerases and, when included in a PCR or other primer-dependent DNA amplification reaction at a concentration, generally not more than 2000 nM, that is effective for at least one of the functions of suppressing mispriming, increasing polymerase selectivity against 3′ terminal mismatches, increasing polymerase selectivity against AT-rich 3′ ends, reducing scatter among replicates, suppressing polymerase 5′ exonuclease activity, and inhibiting polymerase activity; as well as amplification reaction mixtures containing such modified double-stranded oligonucleotides, and amplification reactions, amplification assays and kits that include such modified double-stranded oligonucleotides.

    摘要翻译： 修饰的双链寡核苷酸在其每条链上具有长度为6-50个核苷酸的杂交长度，末端区域具有至少32℃的熔解温度Tm，并且包括2-4个修饰基团， 每个共价连接到不同的末端区域，优选连接到末端核苷酸，所述修饰基团是不具有非平面的庞大部分的多环取代基，所述修饰的寡核苷酸能够结合DNA的5'前核酸酶结构域 聚合酶，并且当包含在浓度通常不大于2000nM的PCR或其它引物依赖性DNA扩增反应中时，对于抑制错配的功能，增加聚合酶对3'末端错配的选择性的至少一个功能是有效的， 增加对富含AT的3'末端的聚合酶选择性，减少重复之间的分散，抑制聚合酶5'核酸外切酶活性和抑制聚合酶a 反应性 以及含有这种修饰的双链寡核苷酸的扩增反应混合物，以及包括这种修饰的双链寡核苷酸的扩增反应，扩增分析和试剂盒。

    Reagents and methods for PCR
    2.
    发明授权
    Reagents and methods for PCR 有权
    试剂和PCR方法

    公开(公告)号：US09034605B2

    公开(公告)日：2015-05-19

    申请号：US13256038

    申请日：2010-03-11

    申请人： Lawrence J. Wangh ,  John Rice ,  Nicholas Rice ,  Yanwei Jia

    发明人： Lawrence J. Wangh ,  John Rice ,  Nicholas Rice ,  Yanwei Jia

    IPC分类号： C12P19/34 ,  C12Q1/68

    CPC分类号： C12Q1/6806 ,  C12Q1/6853 ,  C12Q2545/114 ,  C12Q2531/107 ,  C12Q2527/107

    摘要： Modified double-stranded oligonucleotides that have terminal regions on each of their strands, that have a hybrid length of 6-50 nucleotides long, that have a melting temperature Tm of at least 32° C., and that include 2-4 modifying groups, each covalently attached to a different terminal region, preferably to a terminal nucleotide, said modifying groups being polycyclic substituents that do not have bulky portions that are non-planar, said modified oligonucleotide being capable of binding to the 5′ ex-nuclease domains of DNA polymerases and, when included in a PCR or other primer-dependent DNA amplification reaction at a concentration, generally not more than 2000 nM, that is effective for at least one of the functions of suppressing mispriming, increasing polymerase selectivity against 3′ terminal mismatches, increasing polymerase selectivity against AT-rich 3′ ends, reducing scatter among replicates, suppressing polymerase 5′ exonuclease activity, and inhibiting polymerase activity; as well as amplification reaction mixtures containing such modified double-stranded oligonucleotides, and amplification reactions, amplification assays and kits that include such modified double-stranded oligonucleotides.

    摘要翻译： 修饰的双链寡核苷酸在其每条链上具有长度为6-50个核苷酸的杂交长度，末端区域具有至少32℃的熔解温度Tm，并且包括2-4个修饰基团， 每个共价连接到不同的末端区域，优选连接到末端核苷酸，所述修饰基团是不具有非平面的庞大部分的多环取代基，所述修饰的寡核苷酸能够结合DNA的5'前核酸酶结构域 聚合酶，并且当包含在浓度通常不大于2000nM的PCR或其它引物依赖性DNA扩增反应中时，对于抑制错配的至少一种功能是有效的，增加针对3'末端错配的聚合酶选择性， 提高对富含AT的3'末端的聚合酶选择性，减少重复之间的分散，抑制聚合酶5'核酸外切酶活性，并抑制聚合酶 活动; 以及含有这种修饰的双链寡核苷酸的扩增反应混合物，以及包括这种修饰的双链寡核苷酸的扩增反应，扩增分析和试剂盒。

    METHODS FOR MAKING EMBRYONIC CELLS, EMBRYOS, AND ANIMALS SENSITIZED TO STRESS
    3.
    发明申请
    METHODS FOR MAKING EMBRYONIC CELLS, EMBRYOS, AND ANIMALS SENSITIZED TO STRESS 审中-公开
    制造胚胎细胞，胚胎和敏感的动物的应激方法

    公开(公告)号：US20120198576A1

    公开(公告)日：2012-08-02

    申请号：US13378436

    申请日：2010-07-01

    申请人： Lawrence J. Wangh ,  Cristina Hartshorn ,  Yanwei Jia

    发明人： Lawrence J. Wangh ,  Cristina Hartshorn ,  Yanwei Jia

    IPC分类号： C12N5/0735 ,  A01K67/027 ,  A61K31/343 ,  A61B17/435 ,  A61K33/30 ,  A61K31/198 ,  A61K31/4545 ,  A61K31/7072 ,  C12Q1/68 ,  A61K31/395

    CPC分类号： G01N33/5073 ,  A01K2207/35 ,  C12N15/873

    摘要： Embodiments of the invention are based upon the discovery that exposure of cleavage-stage embryos to a stress inducer, e.g. heat shock or chemical, renders the exposed embryos more sensitive to a secondary treatment with a stress inducer, e.g. heat shock or chemical inducer. Accordingly, the present invention is directed to methods for making embryos, embryonic cells arising from them, and animals and plants that are sensitized to stress, e.g. physiologic or chemical stressors. Methods of screening for inducers and inhibitors of stress using, as test model systems, embryonic cells, embryos, animals, and plants that are sensitized to stress are also disclosed.

    摘要翻译： 本发明的实施方案基于以下发现：切割阶段胚胎暴露于应激诱导物，例如， 热休克或化学，使暴露的胚胎对应激诱导剂的二次治疗更敏感，例如。 热休克或化学诱导剂。 因此，本发明涉及用于制造胚胎，由它们产生的胚胎细胞和致敏致敏的动物和植物的方法，例如， 生理或化学应激物。 还公开了使用致敏应激的作为测试模型系统的胚胎细胞，胚胎，动物和植物来筛选诱导剂和应激抑制剂的方法。

    LATE-PCR
    6.
    发明申请
    LATE-PCR 有权

    公开(公告)号：US20130210656A1

    公开(公告)日：2013-08-15

    申请号：US13734684

    申请日：2013-01-04

    申请人： Lawrence J. Wangh ,  Kenneth Pierce ,  Cristina Hartshorn ,  John Rice ,  J. Aquiles Sanchez

    发明人： Lawrence J. Wangh ,  Kenneth Pierce ,  Cristina Hartshorn ,  John Rice ,  J. Aquiles Sanchez

    IPC分类号： C12Q1/68

    CPC分类号： C12Q1/686 ,  C07H21/00 ,  C12Q1/6806 ,  C12Q1/6851 ,  C12Q2561/113 ,  C12Q2531/107 ,  C12Q2527/107

    摘要： A non-symmetric polymerise chain reaction (PCR) amplification method employing a limiting primer in low concentration whose concentration-adjusted melting point at least equals, and preferably exceeds, that of the excess primer, the latter in turn not being more than 25° C. below the melting temperature of the amplicon. Assays employing such amplification and labeled hybridization probes, including assays that include a detection step following primer extension or a low-temperature probe, or both. Kits for performing such assays and primer or primer-and-probe sets for performing the foregoing amplifications and assays.

    摘要翻译： 使用浓度调节的熔点至少等于并且优选超过过量底漆的极限底物的低浓度限制性引物的非对称聚合链反应（PCR）扩增方法，后者又不超过25℃ 低于扩增子的解链温度。 使用这种扩增和标记的杂交探针的测定法，包括在引物延伸或低温探针之后包括检测步骤的测定法或两者。 用于进行此类测定的试剂盒和用于进行前述扩增和测定的引物或引物 - 探针组。

    Compositions, methods, and kits for detecting and identifying mycobacteria
    8.
    发明授权
    Compositions, methods, and kits for detecting and identifying mycobacteria 有权

    公开(公告)号：US10273525B2

    公开(公告)日：2019-04-30

    申请号：US13884873

    申请日：2011-11-10

    申请人： Lawrence J. Wangh ,  John Rice

    发明人： Lawrence J. Wangh ,  John Rice

    IPC分类号： C07H21/04 ,  C12Q1/6818 ,  C12Q1/689

    摘要： Provided herein are methods for detecting and identifying strains of mycobacteria, and compositions and kits for performing such methods. In particular, nucleic acid amplification and fluorescence detection methods are provided for the detection and differentiation of mycobacteria based on, for example, pathogenicity, species, and antibiotic resistance or sensitivity. Compositions and methods are provided herein to identify and differentiate mycobacteria in mixtures of different mycobacteria and mycobacteria and non-mycobacteria.

    Late-PCR
    9.
    发明授权
    Late-PCR 有权
    晚期PCR

    公开(公告)号：US08367325B2

    公开(公告)日：2013-02-05

    申请号：US11701428

    申请日：2007-02-02

    申请人： Lawrence J. Wangh ,  Kenneth Pierce ,  Cristina Hartshorn ,  John Rice ,  J. Aquiles Sanchez

    发明人： Lawrence J. Wangh ,  Kenneth Pierce ,  Cristina Hartshorn ,  John Rice ,  J. Aquiles Sanchez

    IPC分类号： C12Q1/68 ,  C12P19/34

    CPC分类号： C12Q1/686 ,  C07H21/00 ,  C12Q1/6806 ,  C12Q1/6851 ,  C12Q2561/113 ,  C12Q2531/107 ,  C12Q2527/107

    摘要： A non-symmetric polymerise chain reaction (PCR) amplification method employing a limiting primer in low concentration whose concentration-adjusted melting point at least equals, and preferably exceeds, that of the excess primer, the latter in turn not being more than 25° C. below the melting temperature of the amplicon. Assays employing such amplification and labeled hybridization probes, including assays that include a detection step following primer extension or a low-temperature probe, or both. Kits for performing such assays and primer or primer-and-probe sets for performing the foregoing amplifications and assays.

    摘要翻译： 使用浓度调节的熔点至少等于并且优选超过过量底漆的极限底物的低浓度限制性引物的非对称聚合链反应（PCR）扩增方法，后者又不超过25℃ 低于扩增子的解链温度。 使用这种扩增和标记的杂交探针的测定法，包括在引物延伸或低温探针之后包括检测步骤的测定法或两者。 用于进行此类测定的试剂盒和用于进行前述扩增和测定的引物或引物 - 探针组。