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21. 发明申请

US20080228589A1 Methods For Placing, Accepting, And Filling Orders For Products and Services 审中-公开
标题翻译：用于产品和服务的放置，接受和填写订单的方法
公开(公告)号：US20080228589A1
公开(公告)日：2008-09-18
申请号：US12015143
申请日：2008-01-16
申请人： Ryan T. Koehler , Kenneth J. Livak , Junko Stevens , Francisco M. De La Vega , Michael Rhodes , Laurent R. Bellon , Julie Williams , Dennis A. Gilbert , Charles R. Scafe , Hadar I. Avi-Itzhak , Lily Xu , Susan Eddins , Michael W. Hunkapiller , Leila G. Smith
发明人： Ryan T. Koehler , Kenneth J. Livak , Junko Stevens , Francisco M. De La Vega , Michael Rhodes , Laurent R. Bellon , Julie Williams , Dennis A. Gilbert , Charles R. Scafe , Hadar I. Avi-Itzhak , Lily Xu , Susan Eddins , Michael W. Hunkapiller , Leila G. Smith
IPC分类号： G06Q30/00
CPC分类号： C12Q1/6858 , B01L3/545 , B01L3/5453 , B01L2300/021 , C12Q2600/156 , G01N35/00663 , G06Q30/0621 , G06Q30/0633 , G06Q30/0641 , G06Q50/22 , G16B25/00 , G16B30/00 , G16B35/00 , G16B45/00 , G16B50/00 , G16C20/60 , C12Q2547/101
摘要： Methods and systems for ordering assays which detect SNPs or gene expression are provided. The methods use PCR and RT-PCR procedures. Collections of stock assays are assembled using pre- and post-manufacturing quality control procedures and made available to consumers via the Internet. In addition, custom assays are prepared upon order from the consumer and these assays are also prepared using pre- and post-manufacturing quality control procedures. The assays are then delivered to the consumer.
摘要翻译：提供了检测SNP或基因表达的排序方法和系统。方法采用PCR和RT-PCR方法。库存测定的集合使用制造前和制造后质量控制程序组装，并通过互联网提供给消费者。此外，根据消费者的命令制备定制测定，并且还使用制造前和制造后质量控制程序制备这些测定。然后将测定物递送给消费者。

22. 发明授权

US07414115B2 Length determination of nucleic acid repeat sequences by discontinuous primer extension 有权
标题翻译：通过不连续引物延伸确定核酸重复序列的长度
公开(公告)号：US07414115B2
公开(公告)日：2008-08-19
申请号：US11731192
申请日：2007-03-30
申请人： Kenneth J. Livak , Adam L. Lowe , Andrew J. Blasband
发明人： Kenneth J. Livak , Adam L. Lowe , Andrew J. Blasband
IPC分类号： C07H19/00 , C07H21/02 , C07H21/04 , C12Q1/68 , C12P19/34
CPC分类号： C12Q1/6827 , C12Q1/6837 , C12Q1/6858 , C12Q1/6874 , C12Q2565/514 , C12Q2533/101 , C12Q2525/151 , C12Q2525/131 , C12Q2565/515 , C12Q2525/161 , C12Q2535/125 , C12Q2525/197 , C12Q2563/107
摘要： Disclosed is a kit useful for determining the number of repeat units in a repeat region of a target nucleic acid comprising: a plurality of different-sequence primers, each containing (i) a target binding segment and (ii) a tag segment having a nucleotide sequence that uniquely identifies the target binding segment, a first primer extension reagent; and a second primer extension reagent, wherein at least one of the first or second primer extension reagents includes an extendible nucleotide having a photodestructible label attached thereto.
摘要翻译：公开了一种可用于确定靶核酸重复区域中重复单元数目的试剂盒，其包括：多个不同序列引物，每个引物包含（i）靶结合区段和（ii）具有核苷酸的标签区段唯一识别靶结合片段的序列，第一引物延伸试剂; 和第二引物延伸试剂，其中第一或第二引物延伸试剂中的至少一个包括具有附着于其上的光致可破坏标记的可延伸核苷酸。

23. 发明申请

US20080187913A1 Determination of genotype of an amplification product at multiple allelic sites 审中-公开
标题翻译：确定多个等位基因位点扩增产物的基因型
公开(公告)号：US20080187913A1
公开(公告)日：2008-08-07
申请号：US11592716
申请日：2006-11-03
申请人： Kenneth J. Livak , Federico Goodsaid
发明人： Kenneth J. Livak , Federico Goodsaid
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6858 , C12Q1/6818 , C12Q2561/101 , C12Q2537/143 , C12Q2563/107 , C12Q2531/119
摘要： A method is provided for genotyping a target sequence at least two allelic sites by a 5′ nuclease amplification reaction. In one embodiment, the method includes performing a nucleic acid amplification on a target sequence having at least two different allelic sites using a nucleic acid polymerase having 5′-3′ nuclease activity and a primer capable of hybridizing to the target sequence in the presence of two or more sets of allelic oligonucleotide probes wherein: each set of allelic oligonucleotide probes is for detecting a different allelic site of the target sequence, each set of allelic oligonucleotide probes includes two or more probes which are complementary to different allelic variants at the allelic site being detected by the set of probes, the allelic site being 5′ relative to a sequence to which the primer hybridizes to the target sequence, and at least all but one of the allelic oligonucleotide probes include a different fluorescer than the other probes and a quencher positioned on the probe to quench the fluorescence of the fluorescer; detecting a fluorescence spectrum of the amplification; calculating a fluorescence contribution of each fluorescer to the fluorescence spectrum; and determining a presence or absence of the different allelic variants at the two or more different allelic sites based on the fluorescence contribution of each fluorescer to the combined fluorescence spectrum.
摘要翻译：提供了一种通过5'核酸酶扩增反应将靶序列至少两个等位基因位点进行基因分型的方法。在一个实施方案中，该方法包括使用具有5'-3'核酸酶活性的核酸聚合酶和能够与靶序列杂交的引物在具有至少两个不同等位基因位点的靶序列上进行核酸扩增两组或更多组等位基因寡核苷酸探针，其中：每组等位基因寡核苷酸探针用于检测靶序列的不同等位基因位点，每组等位基因寡核苷酸探针包括与等位基因位点上的不同等位基因变体互补的两个或多个探针被该组探针检测到，等位基因位点相对于引物与靶序列杂交的序列为5'，并且至少所有等位基因寡核苷酸探针中的所有等位基因寡核苷酸探针包括与其它探针不同的荧光剂和猝灭剂位于探针上以淬灭荧光剂的荧光; 检测扩增的荧光光谱; 计算每个荧光剂对荧光光谱的荧光贡献; 并且基于每个荧光剂对组合的荧光光谱的荧光贡献，确定两个或更多个不同等位基因位点处的不同等位基因变体的存在或不存在。

24. 发明授权

US07057025B2 Binary probe and clamp composition 有权
标题翻译：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。

25. 发明授权

US06964848B2 Invasion assay 有权
标题翻译：入侵试验
公开(公告)号：US06964848B2
公开(公告)日：2005-11-15
申请号：US09816150
申请日：2001-03-26
申请人： Kenneth J. Livak , Michael Y. Lucero , Muhammad A. Sharaf
发明人： Kenneth J. Livak , Michael Y. Lucero , Muhammad A. Sharaf
IPC分类号： C12N15/09 , C12Q1/68 , C07H21/02 , C12P19/34
CPC分类号： C12Q1/6823 , C12Q2527/113 , C12Q2537/165 , C12Q2561/109 , C12Q2561/113
摘要： The invention relates to improved methods of detecting and characterizing polynucleotide sequences and variations in polynucleotide sequences. The invention further relates to a device that can be used to detect and characterize polynucleotide sequences and variations in polynucleotide sequences.
摘要翻译：本发明涉及检测和表征多核苷酸序列和多核苷酸序列变异的改进方法。本发明还涉及可用于检测和表征多核苷酸序列和多核苷酸序列变异的装置。

26. 发明授权

US06432642B1 Binary probe and clamp composition and methods for a target hybridization detection 有权
标题翻译：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。

27. 发明授权

US5952202A Methods using exogenous, internal controls and analogue blocks during nucleic acid amplification 失效
标题翻译：在核酸扩增过程中使用外源，内部对照和模拟嵌段的方法
公开(公告)号：US5952202A
公开(公告)日：1999-09-14
申请号：US48880
申请日：1998-03-26
申请人： Kazuko Aoyagi , Kenneth J. Livak
发明人： Kazuko Aoyagi , Kenneth J. Livak
IPC分类号： C07H21/00 , C12Q1/68 , C12P19/34 , C07H21/04
CPC分类号： C07H21/00 , C12Q1/6818
摘要： Reporter-quencher probe assays of nucleic acid amplification, such as PCR, are rendered more meaningful by the addition of internal control reagents. An internal control polynucleotide is amplified with internal control primers and the product is measured by correlation with increased fluorescence by polymerase mediated-exonuclease cleavage or hybridization of the internal control probe. Probes specific for target and internal control polynucleotides are labelled with spectrally resolvable reporters, allowing for concurrent detection and measurement of target and control amplification. A kit of all PCR reagents can be dispensed into reaction chambers in a high-throughput system for rapid and accurate nucleic acid amplification assay, with real-time or end-point measurements. Fluorescent signals correlated to target and internal control levels are spectrally resolvable and measured concurrently. A non-extending oligonucleotide or nucleic analog "block", complementary to the internal control polynucleotide, is added to the amplification mixture to preclude amplification of the internal control polynucleotide and function as an internal negative control. The amplification control reagents, kits, and methods of the present invention provide positive and negative control tests occurring within, and measurable within, the reaction chamber.
摘要翻译：通过添加内部对照试剂使得核酸扩增（例如PCR）的报道 - 猝灭剂探针测定变得更有意义。内部对照多核苷酸用内部对照引物扩增，通过聚合酶介导的外切核酸酶切割或内部对照探针的杂交与荧光增加相关来测量产物。靶向和内部控制多核苷酸的特异性探针用光谱可分辨的报告物标记，允许同时检测和测量靶标和对照扩增。所有PCR试剂的试剂盒可以在高通量系统中分配到反应室中，用于快速准确的核酸扩增测定，具有实时或终点测量。与目标和内部控制水平相关的荧光信号可以光谱解析和并发测量。将与内部对照多核苷酸互补的非延伸寡核苷酸或核苷酸类似物“嵌段”加入到扩增混合物中，以排除内部对照多核苷酸的扩增并作为内部阴性对照起作用。本发明的扩增控制试剂，试剂盒和方法提供在反应室内部和内部可测量的阳性和阴性对照试验。

28. 发明授权

US5736333A Passive internal references for the detection of nucleic acid amplification products 失效
标题翻译：被动内部参考用于检测核酸扩增产物
公开(公告)号：US5736333A
公开(公告)日：1998-04-07
申请号：US657989
申请日：1996-06-04
申请人： Kenneth J. Livak , Lincoln J. McBride
发明人： Kenneth J. Livak , Lincoln J. McBride
IPC分类号： C12N15/09 , C12Q1/68 , C07H21/04 , C12P19/34
CPC分类号： C12Q1/6851
摘要： The invention relates to passive internal references for use in quantitating the formation of amplification products in a nucleic amplification reaction. The internal amplification reference molecules of the invention comprise a first and second fluorophore joined together through a backbone connector. The first and second fluorophores are joined on the backbone in a configuration that permits the energy transfer from the first fluorophore to the second fluorophore. The backbone connector is selected so as not to bind to the target nucleic acid sequence under nucleic acid amplification conditions. Preferably, the backbone connector is a polynucleotide. Another aspect of the invention is to provide passive internal reference molecule containing reagent compositions for use in nucleic acid amplification reactions. The compositions comprise the internal amplification reference molecule of the invention and a nucleic acid amplification reaction buffer. The reagent compositions, optionally, include additional components required for nucleic acid amplification reactions. The invention also provides improved methods of measuring the amount of amplification product in nucleic acid amplification reactions employing fluorescer-quencher probe assays, including methods for the real-time measurement of amplification product formation. The methods comprise the step of adding the internal reference molecule of the invention to the amplification reaction mixture. Fluorescence of the second fluorophore on the internal reference may then be measured and used to calculate changes in fluorescence of the fluorophore on a fluorescer-quencher probe.
摘要翻译：本发明涉及用于量化核酸扩增反应中扩增产物形成的无源内参考。本发明的内部扩增参考分子包含通过骨架连接器连接在一起的第一和第二荧光团。第一和第二荧光团以支持能量从第一荧光团转移到第二荧光团的构型在主链上连接。选择骨架连接器，以便在核酸扩增条件下不与靶核酸序列结合。优选地，骨架连接器是多核苷酸。本发明的另一方面是提供含有用于核酸扩增反应的试剂组合物的被动内参照分子。组合物包含本发明的内部扩增参考分子和核酸扩增反应缓冲液。试剂组合物任选地包括核酸扩增反应所需的另外的组分。本发明还提供了使用荧光猝灭剂探针测定法测量核酸扩增反应中扩增产物量的改进方法，包括用于实时测量扩增产物形成的方法。所述方法包括将本发明的内部参考分子加入到扩增反应混合物中的步骤。然后可以测量第二荧光团在内部参考物上的荧光，并用于计算荧光团在荧光猝灭剂探针上的荧光团的荧光变化。

29. 发明授权

US4879214A Differentiation of nucleic acid segments on the basis of nucleotide differences 失效
标题翻译：基于核苷酸差异分化核酸片段
公开(公告)号：US4879214A
公开(公告)日：1989-11-07
申请号：US272068
申请日：1988-11-15
申请人： John S. Kornher , Kenneth J. Livak
发明人： John S. Kornher , Kenneth J. Livak
IPC分类号： G01N33/58 , C07H21/04 , C12Q1/68
CPC分类号： C12Q1/6858 , C12Q1/6816 , C12Q1/6876 , C12Q2600/156
摘要： A process for distinguishing nucleic acid segments on the basis of nucleotide differences, thereby providing a rapid, convenient means for detecting mutation, is disclosed. The present process comprises synthesizing separately complementary nucleic acid strands on each of at least two target nucleic acid templates using a nucleic acid polymerase and nucleoside triphosphate substrates, wherein at least one of the natural nucleoside triphosphate substrates is replaced with a mobility-shifting analog; denaturing the synthesized strands from the templates if necessary; and comparing the mobility of the separately synthesized strands through a size-fractionation medium. A difference in mobility indicates that the target nucleic acid templates contain different numbers of total nucleotides and/or different numbers of nucleotides complementary to the mobility-shifting analogs. Thus, the process of the present invention can be used to distinguish homologous DNAs or RNAs differing by nucleotide substitutions that affect the number of mobility-shifting analog residues incorporated into synthesized strands, as well as to distinguish differences due to nucleotide insertion or deletion.

30. 发明授权

US08993240B2 Hot start reverse transcription by primer design 有权
标题翻译：通过引物设计热启动逆转录
公开(公告)号：US08993240B2
公开(公告)日：2015-03-31
申请号：US12504633
申请日：2009-07-16
申请人： Kai Qin Lao , Neil A. Straus , Kenneth J. Livak
发明人： Kai Qin Lao , Neil A. Straus , Kenneth J. Livak
IPC分类号： C12Q1/68 , C12P19/34
CPC分类号： C12Q1/6876 , C07H21/02 , C07H21/04 , C12N9/00 , C12Q1/6844 , C12Q1/6848 , C12Q1/686 , C12Q2549/101 , C12Q2533/101 , C12Q2525/301
摘要： The present teachings provide methods, compositions, and kits for performing primer extension reactions. In some embodiments, a reverse transcription reaction is performed on a target polynucleotide with a hot start primer comprising a blunt-ended self-complementary stem, and a loop, and extension products form at high temperatures but reduce extension product formation at low temperatures.
摘要翻译：本教导提供了用于进行引物延伸反应的方法，组合物和试剂盒。在一些实施方案中，对靶多核苷酸进行逆转录反应，所述靶多核苷酸具有包含平端自补偿干杆的热起始引物，并且循环和延伸产物在高温下形成，但是在低温下减少延伸产物形成。

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