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57 条记录 (耗时 0.031 秒)

    Detection method using dissociated rolling circle amplification
    51.
    发明授权
    Detection method using dissociated rolling circle amplification 失效
    使用解离的滚动圆放大的检测方法

    公开(公告)号：US07553619B2

    公开(公告)日：2009-06-30

    申请号：US10072666

    申请日：2002-02-08

    申请人： Gyanendra Kumar ,  Patricio Abarzua ,  Michael Egholm

    发明人： Gyanendra Kumar ,  Patricio Abarzua ,  Michael Egholm

    IPC分类号： C12Q1/68 ,  C07H21/04

    CPC分类号： G01N33/537 ,  C12Q1/6804 ,  C12Q2563/179 ,  C12Q2531/125

    摘要： Disclosed are compositions and methods for detecting small quantities of analytes such as proteins and peptides. The method involves associating a DNA circle with the analyte and subsequent release and rolling circle replication of the circular DNA molecule. In the method, an amplification target circle is associated with analytes using a conjugate of the circle and a specific binding molecule that is specific for the analyte to be detected. Amplification target circles not associated with the proteins are removed, the amplification target circles that are associated with the proteins are decoupled from the specific binding molecule and amplified by rolling circle amplification. The amplification is isothermic and can result in the production of a large amount of nucleic acid from each primer. The amplified DNA serves as a readily detectable signal for the analytes.

    摘要翻译： 公开了用于检测少量分析物如蛋白质和肽的组合物和方法。 该方法包括将DNA环与分析物结合，并随后循环DNA分子的释放和循环复制。 在该方法中，扩增目标圆与分析物相关联，所述分析物使用圆的缀合物和对待检测分析物特异的特异性结合分子。 去除与蛋白质不相关的扩增靶区，与蛋白质相关的扩增靶区与特异性结合分子解偶联并通过滚环扩增进行扩增。 扩增是等温的，并且可以导致从每个引物产生大量的核酸。 扩增的DNA作为分析物的易检测信号。

    BINARY PROBE AND CLAMP COMPOSITION AND METHODS FOR TARGET HYBRIDIZATION DETECTION
    52.
    发明申请
    BINARY PROBE AND CLAMP COMPOSITION AND METHODS FOR TARGET HYBRIDIZATION DETECTION 审中-公开
    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：US20070026429A1

    公开(公告)日：2007-02-01

    申请号：US11422211

    申请日：2006-06-05

    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。

    Polymerase extension at 3' terminus of PNA-DNA chimera
    53.
    发明申请
    Polymerase extension at 3' terminus of PNA-DNA chimera 审中-公开
    PNA-DNA嵌合体3'末端的聚合酶延伸

    公开(公告)号：US20050186572A1

    公开(公告)日：2005-08-25

    申请号：US10618077

    申请日：2003-07-11

    申请人： Michael Egholm ,  Caifu Chen

    发明人： Michael Egholm ,  Caifu Chen

    IPC分类号： G01N33/53 ,  C12N15/09 ,  C12Q1/68 ,  G01N21/78 ,  G01N33/566 ,  G01N33/58 ,  C07K14/00

    CPC分类号： C12Q1/6858 ,  C12Q1/6853 ,  C12Q1/6869 ,  C12Q2525/107 ,  C12Q2537/143

    摘要： The invention provides methods and kits for primer extension of PNA-DNA chimera from template nucleic acids using polymerases, nucleotide 5′-triphosphates, and primer extension reagents. Structural requirements of the chimera for primer extension include 5 to 15 contiguous PNA monomer units, 3 or more contiguous nucleotides, and a 3′ hydroxyl terminus. The chimera and/or a nucleotide is labelled with fluorescent dyes or other labels. The methods include DNA sequencing, DNA fragment analysis, reverse transcription, mini-sequencing, chromosome labelling, amplification, and single nucleotide polymorphism (SNP) detection.

    摘要翻译： 本发明提供了使用聚合酶，核苷酸5'-三磷酸和引物延伸试剂从模板核酸引物延伸PNA-DNA嵌合体的方法和试剂盒。 用于引物延伸的嵌合体的结构要求包括5至15个连续的PNA单体单元，3个或更多个连续核苷酸和3'羟基末端。 嵌合体和/或核苷酸用荧光染料或其他标记物标记。 方法包括DNA测序，DNA片段分析，逆转录，微型测序，染色体标记，扩增和单核苷酸多态性（SNP）检测。

    Asynchronous primed PCR
    54.
    发明申请
    Asynchronous primed PCR 审中-公开
    异步引物PCR

    公开(公告)号：US20050130178A1

    公开(公告)日：2005-06-16

    申请号：US10865683

    申请日：2004-06-09

    申请人： Caifu Chen ,  Michael Egholm ,  Lawrence Haff

    发明人： Caifu Chen ,  Michael Egholm ,  Lawrence Haff

    IPC分类号： G01N33/53 ,  C12N15/09 ,  C12Q1/68 ,  G01N33/566 ,  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.

    摘要翻译： 用于核酸扩增的异步热循环方案使用两种引物，其热解温度不同于约10至30℃。在较高熔点引物退火和聚合酶介导的延伸之后，未接合的单链靶序列可能被杂交和检测 通过探针。 DNA探针可能被聚合酶的外切核酸酶活性切割。 探针可以是非断裂的类似物，例如PNA。 当探针用报告染料和选择进行能量转移的猝灭剂标记时，例如， 当探针未结合时，来自报告染料的荧光可以被有效地淬灭。 当探针与互补靶物杂交时或在与靶标结合时进行切割时，报道染料不再猝灭，导致可检测量的荧光。 可以将第二种较低熔点的引物退火并延伸以产生双链核酸。 可以实时监测放大，包括每个周期，或在终点。 异构PCR热循环方案可以在退火和延伸较高熔点引物的方案结束时通过重复生成单链形式的PCR扩增子的优势。

    Methods and kit for hybridization anaylsis using peptide nucleic acid probes
    55.
    发明申请
    Methods and kit for hybridization anaylsis using peptide nucleic acid probes 审中-公开
    使用肽核酸探针进行杂交分析的方法和试剂盒

    公开(公告)号：US20050053944A1

    公开(公告)日：2005-03-10

    申请号：US10636343

    申请日：2003-08-07

    申请人： Martin Fuchs ,  Michael Egholm ,  Heather O'Keefe ,  Xian-Wei Yao

    发明人： Martin Fuchs ,  Michael Egholm ,  Heather O'Keefe ,  Xian-Wei Yao

    IPC分类号： G01N33/53 ,  C12N15/09 ,  C12Q1/68 ,  C12Q1/6813 ,  G01N27/447 ,  G01N33/566 ,  C07K14/47

    CPC分类号： G01N27/44726 ,  C12Q1/6813 ,  G01N27/447 ,  C12Q2525/113 ,  C12Q2523/113 ,  C12Q2565/125 ,  C12Q2565/629 ,  C12Q2565/107

    摘要： A method composition and apparatus for the hybridization and separation of molecules having a desired target sequence in a sample by contacting a sample of single stranded nucleic acids with a detectable PNA probe having a sequence complementary to the target sequence so that the target sequence, if present, will hybridize with the detectable probe to form a detectable duplex, and then separating the duplex in a denaturing medium from unbound sample components by electrophoresis. The invention also relates to methods compositions and apparatus for the hybridization and separation of molecules having a desired target sequence in a mixed sample of single stranded nucleic acids and their complementary strands by contacting the sample with a detectable PNA probe.

    摘要翻译： 一种方法组合物和装置，用于通过使单链核酸样品与具有与靶序列互补的序列的可检测PNA探针接触来杂交和分离样品中具有所需靶序列的分子，使得靶序列（如果存在） 将与可检测的探针杂交以形成可检测的双链体，然后通过电泳将变性培养基中的双链体与未结合的样品组分分离。 本发明还涉及用于通过使样品与可检测的PNA探针接触来在单链核酸及其互补链的混合样品中杂交和分离具有所需靶序列的分子的方法组合物和装置。