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    Template-dependent ligation with PNA-DNA chimeric probes
    43.
    发明授权
    Template-dependent ligation with PNA-DNA chimeric probes 有权
    与PNA-DNA嵌合探针的模板依赖性连接

    公开(公告)号：US06297016B1

    公开(公告)日：2001-10-02

    申请号：US09416003

    申请日：1999-10-08

    申请人： Michael Egholm ,  Caifu Chen

    发明人： Michael Egholm ,  Caifu Chen

    IPC分类号： C12Q168

    CPC分类号： C12Q1/6862

    摘要： The invention provides methods, kits, and compositions for ligation of PNA-DNA chimeric probes and oligonucleotides when they are hybridized adjacently to template nucleic acids using ligases and ligation reagents. Structural requirements of the chimeras for ligation include 5 to 15 contiguous PNA monomer units, 2 or more contiguous nucleotides, and a 3′ hydroxyl or 5′ hydroxyl terminus. The chimera and/or oligonucleotide may be labelled with fluorescent dyes or other labels. The methods include, for example, oligonucleotide-ligation assays (OLA) and single nucleotide polymorphism detection.

    摘要翻译： 当使用连接酶和连接试剂将它们与模板核酸相互杂交时，本发明提供用于连接PNA-DNA嵌合探针和寡核苷酸的方法，试剂盒和组合物。 用于连接的嵌合体的结构要求包括5至15个连续的PNA单体单元，2个或更多个连续核苷酸，以及3'羟基或5'羟基末端。 嵌合体和/或寡核苷酸可以用荧光染料或其他标记物标记。 所述方法包括例如寡核苷酸连接测定（OLA）和单核苷酸多态性检测。

    Synthons for the synthesis and deprotection of peptide nucleic acids under mild conditions
    44.
    发明授权
    Synthons for the synthesis and deprotection of peptide nucleic acids under mild conditions 失效
    用于在温和条件下合成和去保护肽核酸的合成子

    公开(公告)号：US6133444A

    公开(公告)日：2000-10-17

    申请号：US487666

    申请日：1995-06-07

    申请人： James M. Coull ,  Michael Egholm ,  Richard P. Hodge ,  Mohamed Ismail ,  Sharanappa B. Rajur

    发明人： James M. Coull ,  Michael Egholm ,  Richard P. Hodge ,  Mohamed Ismail ,  Sharanappa B. Rajur

    IPC分类号： C07D239/47 ,  C07D239/46 ,  C07D473/18 ,  C07D473/34 ,  C07D473/40 ,  C07K14/00

    CPC分类号： C07D239/47 ,  C07D473/18 ,  C07D473/34 ,  C07D473/40 ,  C07K14/003 ,  Y02P20/55

    摘要： A method is disclosed for preparing novel purine PNA synthons having protecting groups which may be removed under mild conditions. The purine PNA synthons generally are prepared by coupling purine derivatives having carbamate protection to a protected N-(2-aminoethyl)-glycine backbone. By a method of this invention, purine PNA synthons may have orthogonal protection of the carbamate protected purine and the protected backbone. The purine PNA synthons are useful in the synthesis of peptide nucleic acids (PNAs) and other oligomers such as PNA-DNA chimeras, and may be used in automated synthesizers. In practicing methods of the invention, novel compositions of matter also are disclosed. For example, disclosed herein are an adenine PNA synthon having the following formula: ##STR1## and a guanine PNA synthon having the following formula: ##STR2##

    摘要翻译： 公开了一种制备具有可在温和条件下除去的保护基的新型嘌呤PNA合成子的方法。 嘌呤PNA合成酶通常通过将具有氨基甲酸酯保护的嘌呤衍生物与保护的N-（2-氨基乙基） - 甘氨酸主链偶联来制备。 通过本发明的方法，嘌呤PNA合成子可具有氨基甲酸酯保护的嘌呤和被保护的主链的正交保护。 嘌呤PNA合成子可用于合成肽核酸（PNA）和其它寡聚体如PNA-DNA嵌合体，并可用于自动合成仪中。 在本发明的实践方法中，还公开了新的物质组合物。 例如，本文公开了具有下式的腺嘌呤PNA合成子和具有下式的鸟嘌呤PNA合成子：

    BINARY PROBE AND CLAMP COMPOSITION AND METHODS FOR TARGET HYBRIDIZATION DETECTION
    45.
    发明申请
    BINARY PROBE AND CLAMP COMPOSITION AND METHODS FOR TARGET HYBRIDIZATION DETECTION 有权
    二次探针和夹钳组合物和方法进行目标杂交检测

    公开(公告)号：US20100291557A1

    公开(公告)日：2010-11-18

    申请号：US12538823

    申请日：2009-08-10

    申请人： Kenneth J. Livak ,  Michael Egholm ,  Michael W. Hunkapiller

    发明人： Kenneth J. Livak ,  Michael Egholm ,  Michael W. Hunkapiller

    IPC分类号： C12Q1/68 ,  C07H21/04

    CPC分类号： C12Q1/6816 ,  C12Q1/6818 ,  C12Q2549/126 ,  C12Q2537/119 ,  C12Q2525/179

    摘要： Binary probe and clamp compositions conduct methods for target hybridization detection. Where the probe is a substrate for exonuclease cleavage, the composition provides quantitation and detection of PCR products, by real-time and end-point measurements. Where the probe is an amplification primer, the composition provides an improved method for labelling and detection of PCR products. Probes and clamps may be labelled with fluorescent dyes, quenchers, hybridization-stabilizing moieties, chemiluminescent dyes, and affinity ligands. Clamps may be nucleic acid analogs, such as 2-aminoethylglycine PNA.

    摘要翻译： 二进制探针和夹具组合物进行靶杂交检测的方法。 当探针是外切核酸酶切割的底物时，组合物通过实时和终点测量提供PCR产物的定量和检测。 当探针是扩增引物时，组合物提供了PCR产物的标记和检测的改进方法。 探针和夹具可以用荧光染料，猝灭剂，杂交稳定部分，化学发光染料和亲和配体标记。 夹具可以是核酸类似物，例如2-氨基乙基甘氨酸PNA。

    Binary probe and clamp composition
    46.
    发明授权
    Binary probe and clamp composition 有权
    二进制探针和夹具组成

    公开(公告)号：US07057025B2

    公开(公告)日：2006-06-06

    申请号：US10112677

    申请日：2002-03-28

    申请人： Kenneth J. Livak ,  Michael Egholm ,  Michael W. Hunkapiller

    发明人： Kenneth J. Livak ,  Michael Egholm ,  Michael W. Hunkapiller

    IPC分类号： C07H21/02 ,  C07H21/04 ,  C12Q1/68 ,  C12P19/34

    CPC分类号： C12Q1/6816 ,  C12Q1/6818 ,  C12Q2549/126 ,  C12Q2537/119 ,  C12Q2525/179

    摘要： Binary probe and clamp compositions conduct methods for target hybridization detection. Where the probe is a substrate for exonuclease cleavage, the composition provides quantitation and detection of PCR products, by real-time and end-point measurements. Where the probe is an amplification primer, the composition provides an improved method for labelling and detection of PCR products. Probes and clamps may be labelled with fluorescent dyes, quenchers, hybridization-stabilizing moieties, chemiluminescent dyes, and affinity ligands. Clamps may be nucleic acid analogs, such as 2-aminoethylglycine PNA.

    摘要翻译： 二进制探针和夹具组合物进行靶杂交检测的方法。 当探针是外切核酸酶切割的底物时，组合物通过实时和终点测量提供PCR产物的定量和检测。 当探针是扩增引物时，组合物提供了PCR产物的标记和检测的改进方法。 探针和夹具可以用荧光染料，猝灭剂，杂交稳定部分，化学发光染料和亲和配体标记。 夹具可以是核酸类似物，例如2-氨基乙基甘氨酸PNA。

    BINARY PROBE AND CLAMP COMPOSITION.
    47.
    发明申请
    BINARY PROBE AND CLAMP COMPOSITION. 有权
    二进制和夹子组成。

    公开(公告)号：US20060035217A1

    公开(公告)日：2006-02-16

    申请号：US10112677

    申请日：2002-03-28

    申请人： Kenneth Livak ,  Michael Egholm ,  Michael Hunkapiller

    发明人： Kenneth Livak ,  Michael Egholm ,  Michael Hunkapiller

    IPC分类号： C12Q1/68 ,  C07H21/04

    CPC分类号： C12Q1/6816 ,  C12Q1/6818 ,  C12Q2549/126 ,  C12Q2537/119 ,  C12Q2525/179

    摘要： Binary probe and clamp compositions conduct methods for target hybridization detection. Where the probe is a substrate for exonuclease cleavage, the composition provides quantitation and detection of PCR products, by real-time and end-point measurements. Where the probe is an amplification primer, the composition provides an improved method for labelling and detection of PCR products. Probes and clamps may be labelled with fluorescent dyes, quenchers, hybridization-stabilizing moieties, chemiluminescent dyes, and affinity ligands. Clamps may be nucleic acid analogs, such as 2-aminoethylglycine PNA.

    摘要翻译： 二进制探针和夹具组合物进行靶杂交检测的方法。 当探针是外切核酸酶切割的底物时，组合物通过实时和终点测量提供PCR产物的定量和检测。 当探针是扩增引物时，组合物提供了PCR产物的标记和检测的改进方法。 探针和夹具可以用荧光染料，猝灭剂，杂交稳定部分，化学发光染料和亲和配体标记。 夹具可以是核酸类似物，例如2-氨基乙基甘氨酸PNA。

    Asynchronous primed PCR
    48.
    发明授权
    Asynchronous primed PCR 有权

    公开(公告)号：US06887664B2

    公开(公告)日：2005-05-03

    申请号：US09875211

    申请日：2001-06-05

    申请人： Caifu Chen ,  Michael Egholm ,  Lawrence A. Haff

    发明人： Caifu Chen ,  Michael Egholm ,  Lawrence A. Haff

    IPC分类号： G01N33/53 ,  C12N15/09 ,  C12Q1/68 ,  G01N33/566 ,  C07H21/00 ,  C07H21/02 ,  C12P19/34

    CPC分类号： C12Q1/686 ,  C12Q2561/101 ,  C12Q2527/113 ,  C12Q2527/107

    摘要： An asynchronous thermal cycling protocol for nucleic acid amplification uses two primers with thermal melting temperatures different by about 10 to 30° C. After the higher melting primer has annealed and polymerase mediated extension, the uncopied, single-stranded target sequence may be hybridized and detected by a probe. DNA probes may be cleaved by the exonuclease activity of a polymerase. The probe may be a non-cleaving analog such as PNA. When a probe is labelled with a reporter dye and a quencher selected to undergo energy transfer, e.g. FRET, fluorescence from the reporter dye may be effectively quenched when the probe is unbound. Upon hybridization of the probe to complementary target or upon cleavage while bound to target, the reporter dye is no longer quenched, resulting in a detectable amount of fluorescence. The second, lower-melting primer may be annealed and extended to generate a double-stranded nucleic acid. Amplification may be monitored in real time, including each cycle, or at the end point. The asynchronous PCR thermal cycling protocol can generate a preponderance of the PCR amplicon in single-stranded form by repetition at the end of the protocol of annealing and extension of the higher melting primer.

    Binary probe and clamp composition and methods for a target hybridization detection
    49.
    发明授权
    Binary probe and clamp composition and methods for a target hybridization detection 有权
    用于靶杂交检测的二进制探针和夹钳组成及方法

    公开(公告)号：US06432642B1

    公开(公告)日：2002-08-13

    申请号：US09232000

    申请日：1999-01-15

    申请人： Kenneth J. Livak ,  Michael Egholm ,  Michael W. Hunkapiller

    发明人： Kenneth J. Livak ,  Michael Egholm ,  Michael W. Hunkapiller

    IPC分类号： C12Q168

    CPC分类号： C12Q1/6816 ,  C12Q1/6818 ,  C12Q2549/126 ,  C12Q2537/119 ,  C12Q2525/179

    摘要： Binary probe and clamp compositions conduct methods for target hybridization detection. Where the probe is a substrate for exonuclease cleavage, the composition provides quantitation and detection of PCR products, by real-time and end-point measurements. Where the probe is an amplification primer, the composition provides an improved method for labelling and detection of PCR products. Probes and clamps may be labelled with fluorescent dyes, quenchers, hybridization-stabilizing moieties, chemiluminescent dyes, and affinity ligands. Clamps may be nucleic acid analogs, such as 2-aminoethylglycine PNA.

    摘要翻译： 二进制探针和夹具组合物进行靶杂交检测的方法。 当探针是外切核酸酶切割的底物时，组合物通过实时和终点测量提供PCR产物的定量和检测。 当探针是扩增引物时，组合物提供了PCR产物的标记和检测的改进方法。 探针和夹具可以用荧光染料，猝灭剂，杂交稳定部分，化学发光染料和亲和配体标记。 夹具可以是核酸类似物，例如2-氨基乙基甘氨酸PNA。

    Nucleic acid amplification
    50.
    发明授权
    Nucleic acid amplification 有权
    核酸扩增

    公开(公告)号：US09487823B2

    公开(公告)日：2016-11-08

    申请号：US10327602

    申请日：2002-12-20

    申请人： Roger S. Lasken ,  Michael Egholm ,  Osama A. Alsmadi

    发明人： Roger S. Lasken ,  Michael Egholm ,  Osama A. Alsmadi

    IPC分类号： C12Q1/68

    CPC分类号： C12Q1/6844 ,  C12Q1/6858 ,  C12Q2531/119 ,  C12Q2525/151

    摘要： Disclosed are compositions and a method for amplification of nucleic acid sequences of interest. The disclosed method generally involves replication of a complex nucleic acid sample such as genomic samples using one, a few, or more primers such that, during replication, the replicated strands are displaced from the nucleic acid molecules in the sample by strand displacement replication of another replicated strand. It was discovered that highly complex nucleic acid samples can be efficiently amplified using only one or a few primers having specific nucleic acid sequences. The one or few primers are complementary to nucleic acid sequences distributed throughout nucleic acid in the sample.

    摘要翻译： 公开了用于扩增感兴趣的核酸序列的组合物和方法。 所公开的方法通常涉及使用一种，几种或多种引物复制复杂核酸样品，例如基因组样品，使得在复制期间，复制的链通过另一个的链置换复制从样品中的核酸分子中移位 复制链。 已经发现，仅使用具有特定核酸序列的一个或几个引物可以有效扩增高度复杂的核酸样品。 一个或几个引物与分析在样品中的核酸的核酸序列互补。