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1. 发明申请

US20100112557A1 METHOD FOR HIGH RESOLUTION MELT GENOTYPING 审中-公开
标题翻译：高分辨率熔融基因分析方法
公开(公告)号：US20100112557A1
公开(公告)日：2010-05-06
申请号：US12264193
申请日：2008-11-03
申请人： Andreas R. TOBLER
发明人： Andreas R. TOBLER
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6848 , C12Q1/6858 , C12Q1/686 , C12Q2565/301 , C12Q2525/186 , C12Q2521/101 , C12Q2527/107
摘要： Various methods are described that provide for high resolution melt (HRM) genotyping. The embodiments include providing a locus specific primer and two allele specific primers each having a 5′ end with a short tail, providing a nucleic acid having a single nucleotide polymorphism (SNP) base located within 1-20 base pairs of the 3′ end of nucleic acid, hybridizing the locus specific primer and the allele specific primers to the nucleic acid, amplifying the sample using pyrophosphorolysis activated polymerization (PAP) PCR enzyme, and determining the Tm of the amplicons using HRM. In other embodiments, reactions mixtures and kits for HRM genotyping are provided and disclosed. These kits comprise a locus specific primer, one or more allele specific primers each having a 5′ end with a short tail, a nucleic acid, and a pyrophosphorolysis activate polymerization (PAP) PCR enzyme.
摘要翻译：描述了提供高分辨率熔体（HRM）基因分型的各种方法。实施方案包括提供基因座特异性引物和两个等位基因特异性引物，每个具有5'末端具有短尾巴，提供具有单核苷酸多态性（SNP）碱基的核苷酸，其位于3'端的1' 核酸，将基因座特异性引物和等位基因特异性引物杂交到核酸，使用焦磷酸裂解活化聚合（PAP）PCR酶扩增样品，并使用HRM测定扩增子的Tm。在其它实施方案中，提供并披露用于HRM基因分型的反应混合物和试剂盒。这些试剂盒包含一个基因座特异性引物，一个或多个等位基因特异性引物，每个具有5'末端具有短尾巴，核酸和焦磷酸裂解活化聚合（PAP）PCR酶。

2. 发明申请

US20090209008A1 THERMUS THERMOPHILUS NUCLEIC ACID POLYMERASES 审中-公开
标题翻译： THERMUS THERMOPHILUS核酸聚合物
公开(公告)号：US20090209008A1
公开(公告)日：2009-08-20
申请号：US12193691
申请日：2008-08-18
申请人： JAMES E. ROZZELLE , ELENA V. BOLCHAKOVA
发明人： JAMES E. ROZZELLE , ELENA V. BOLCHAKOVA
IPC分类号： C12P19/34
CPC分类号： C12N9/1252 , C12P19/34 , C12Q1/686 , C12Y207/07007 , Y10S435/975
摘要： The invention provides novel nucleic acid polymerases from strains GK24 and RQ-1 of Thermus thermophilus, and nucleic acids encoding those polymerases, as well as methods for using the polymerases and nucleic acids.
摘要翻译：本发明提供了嗜热栖热菌菌株GK24和RQ-1的新型核酸聚合酶和编码这些聚合酶的核酸，以及使用聚合酶和核酸的方法。

3. 发明申请

US20090191553A1 Chase Ligation Sequencing 审中-公开
标题翻译：追逐连锁测序
公开(公告)号：US20090191553A1
公开(公告)日：2009-07-30
申请号：US12243925
申请日：2008-10-01
申请人： Cynthia Hendrickson
发明人： Cynthia Hendrickson
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6874 , C12Q2537/155 , C12Q2533/107 , C12Q2525/161 , C12Q2525/185
摘要： In various embodiments, the present teachings provide sequencing methods which facilitate enhancing the efficiency of ligation and/or increasing sequencing reads. Various embodiments of the methods enable sequencing through template regions for which complementary labeled extension probes are unavailable or insufficient. In various embodiments, one or more rounds of ligation with unlabeled extension probes can be used in addition to a round of ligation with labeled extension probe. In various embodiments, for example, such methods can facilitate extension on template polynucleotides that do not bind labeled extension probe in the first round of ligation.
摘要翻译：在各种实施方案中，本教导提供了有助于提高连接效率和/或增加测序读数的测序方法。这些方法的各种实施方案使得能够通过补体标记的扩增探针不可用或不足的模板区进行测序。在各种实施方案中，除了用标记的延伸探针的一轮连接之外，还可以使用与未标记的延伸探针的一轮或多轮连接。在各种实施方案中，例如，这样的方法可以有助于在第一轮连接中不结合标记的延伸探针的模板多核苷酸上的延伸。

4. 发明申请

US20090181860A1 Reagents, methods, and libraries for bead-based sequencing 有权
标题翻译：用于珠基测序的试剂，方法和文库
公开(公告)号：US20090181860A1
公开(公告)日：2009-07-16
申请号：US12220208
申请日：2008-07-21
申请人： Kevin McKernan , Alan Blanchard , Lev Kotler , Gina Costa
发明人： Kevin McKernan , Alan Blanchard , Lev Kotler , Gina Costa
IPC分类号： C40B40/06 , C07H21/04
CPC分类号： C12Q1/6874 , B82Y15/00 , B82Y30/00 , C12Q1/68 , C12Q1/6837 , C12Q1/6844 , C12Q1/6869 , C12Q2565/537 , C12Q2565/518 , C12Q2565/513 , C12Q2565/137 , C12Q2565/102 , C12Q2533/107 , C12Q2537/165
摘要： The present invention provides methods for determining a nucleic acid sequence by performing successive cycles of duplex extension along a single stranded template. The cycles comprise steps of extension, ligation, and, preferably, cleavage. In certain embodiments the methods make use of extension probes containing phosphorothiolate linkages and employ agents appropriate to cleave such linkages. The invention provides methods of determining information about a sequence using at least two distinguishably labeled probe families. In certain embodiments the methods acquire less than 2 bits of information from each of a plurality of nucleotides in the template in each cycle. In certain embodiments the sequencing reactions are performed on templates attached to immobilized beads. The invention further provides sets of labeled probes containing phosphorothiolate linkages. In addition, the invention includes performing multiple sequencing reactions on a single template by removing initializing oligonucleotides and extended strands and performing subsequent reactions using different initializing oligonucleotides.
摘要翻译：本发明提供了通过沿着单链模板进行连续循环的双链延伸来确定核酸序列的方法。循环包括延伸，连接和优选切割的步骤。在某些实施方案中，所述方法利用含有硫代磷酸酯键的延伸探针，并使用适合切割这种连接的试剂。本发明提供使用至少两个可区分标记的探针家族确定关于序列的信息的方法。在某些实施方案中，该方法在每个周期中从模板中的多个核苷酸中的每一个获取少于2位的信息。在某些实施方案中，测序反应在附着于固定化珠粒的模板上进行。本发明还提供了含有硫代磷酸酯键的标记探针的集合。此外，本发明包括通过除去初始化寡核苷酸和延伸的链并使用不同的初始化寡核苷酸进行后续反应，在单个模板上进行多个测序反应。

5. 发明申请

US20090171078A1 METHOD OF SEQUENCING NUCLEIC ACIDS USING ELABORATED NUCLEOTIDE PHOSPHOROTIOLATE COMPOUNDS 审中-公开
标题翻译：使用制备的核酸磷酸酯化合物测序核酸的方法
公开(公告)号：US20090171078A1
公开(公告)日：2009-07-02
申请号：US12275176
申请日：2008-11-20
申请人： Kai Qin LAO , Neil A. Straus
发明人： Kai Qin LAO , Neil A. Straus
IPC分类号： C07H19/04 , C12P19/34 , C12Q1/68
CPC分类号： C12Q1/6883
摘要： The present teachings provide methods, compositions, and kits for synthesizing and sequencing nucleic acids. In some embodiments, elaborated nucleotide phosphorothiolate compounds are employed along with efficient cleaving reactions. Improved sequencing efficiency is achieved by the rapid polymerase-mediated incorporation of elaborated nucleotide phosphorothiolate compounds. Increased sequencing efficiency is also achieved by the ability of the cleaving reactions to restore the incorporated nucleotides to their natural structure prior to subsequent elongation.
摘要翻译：本教导提供了用于合成和测序核酸的方法，组合物和试剂盒。在一些实施方案中，精制的核苷酸硫代磷酸酯化合物与有效的切割反应一起使用。通过快速聚合酶介导的拟合的核苷酸硫代硫酸酯化合物的掺入可以改善测序效率。增加的测序效率也可以通过切割反应在随后的伸长之前将掺入的核苷酸恢复到其天然结构的能力来实现。

6. 发明申请

US20090099027A1 Methods of Modifying Support Surfaces for the Immobilization of Particles and the Use of the Immobilized Particles for Analyzing Nucleic Acids 审中-公开
标题翻译：用于固定颗粒的支撑表面的修饰方法和固定化颗粒用于分析核酸的用途
公开(公告)号：US20090099027A1
公开(公告)日：2009-04-16
申请号：US12197132
申请日：2008-08-22
申请人： Douglas P. GREINER , Jeremy STUART , Meng C. TAING , Steven M. MENCHEN
发明人： Douglas P. GREINER , Jeremy STUART , Meng C. TAING , Steven M. MENCHEN
IPC分类号： C40B20/04 , C40B50/18 , C40B40/04
CPC分类号： C40B20/04 , C07B2200/11 , C07C335/20 , C07H21/00 , C12Q1/6869 , C40B40/04 , C40B40/06 , C40B40/08 , C40B50/18 , C40B80/00 , C12Q2565/518 , C12Q2521/501 , C12Q2523/107 , C12Q2533/101
摘要： Methods of modifying a nucleophilic surface of a support are described. The methods involve reacting a multifunctional electrophilic reagent with nucleophilic groups on the surface of the support. The resulting electrophilic surface can be used for the covalent attachment of particles (e.g. beads) having nucleophilic functional groups. For example, nucleic acid templates with nucleophilic (e.g., amine) groups can be attached to a surface of the particles. The nucleophilic groups on the nucleic acid templates can then be used to attach the particles to the modified surface of the support. The resulting support-bound particles can be used to analyze (e.g., sequence) the nucleic acid templates on the particles.
摘要翻译：描述了改变载体的亲核表面的方法。所述方法包括使多功能亲电试剂与载体表面上的亲核基团反应。所得的亲电表面可以用于具有亲核官能团的颗粒（例如珠粒）的共价连接。例如，具有亲核（例如胺）基团的核酸模板可以附着于颗粒的表面。然后核酸模板上的亲核基团可用于将颗粒附着到载体的改性表面上。得到的载体结合颗粒可用于分析（例如，序列）颗粒上的核酸模板。

7. 发明授权

US07491830B2 Phenyl xanthene dyes 有权
标题翻译：苯基呫吨染料
公开(公告)号：US07491830B2
公开(公告)日：2009-02-17
申请号：US10837621
申请日：2004-05-04
申请人： Joe Y. L. Lam , Steven M. Menchen , Ruiming Zou , Scott C. Benson
发明人： Joe Y. L. Lam , Steven M. Menchen , Ruiming Zou , Scott C. Benson
IPC分类号： C09B11/24
CPC分类号： C09B11/24 , C07D311/82 , C07D491/14 , C09B11/06 , C09B11/12 , C09B11/22 , C09B11/26 , C09B13/00 , C09B57/00 , C09B62/0025
摘要： Fluorescent phenyl xanthene dyes are described that comprise any fluorescein, rhodamine or rhodol comprising a particular C9 phenyl ring. One or both of the ortho groups on the lower C9 phenyl ring is ortho substituted with a group selected from alkyl, heteroalkyl, alkoxy, halo, haloalkyl, amino, mercapto, alkylthio, cyano, isocyano, cyanato, mercaptocyanato, nitroso, nitro, azido, sulfeno, sulfinyl, and sulfino. In one embodiment, halo and/or hydroxy groups are used. Optimal dyes contain a lower C9 phenyl ring in which both ortho groups are the same and the lower ring exhibits some form a symmetry relative to an imaginary axis running from the phenyl rings point of attachment to the remainder of the xanthene dye through a point para to the point of attachment. The phenyl xanthene dyes may be activated. Furthermore, the phenyl xanthene dyes may be conjugated to one or more substances including other dyes. The phenyl xanthene dyes are useful for a number of purposes, including labels for use in automated DNA sequencing as well the formation of fluorescent “bar codes” for polymeric particles used in the multiplexed analysis of analytes.
摘要翻译：描述了包含任何含有特定C 9苯环的任何荧光素，罗丹明或若丹明的荧光苯基呫吨染料。下C9苯环上的一个或两个邻基被选自烷基，杂烷基，烷氧基，卤素，卤代烷基，氨基，巯基，烷硫基，氰基，异氰基，氰基，巯基氰基，亚硝基，硝基，叠氮基，磺酰亚胺基，亚磺酰基和亚磺酰基。在一个实施方案中，使用卤素和/或羟基。最佳染料含有较低的C 9苯环，其中两个邻基团相同，并且下环表现出相对于从苯环连接点到呫吨染料的其余部分的假想轴的对称性。附件点。苯基呫吨染料可以被活化。此外，苯基呫吨染料可以与一种或多种包括其它染料的物质共轭。苯基呫吨染料可用于许多目的，包括用于自动DNA测序的标记，以及用于分析物复用分析中使用的聚合物颗粒的荧光“条形码”。

8. 发明申请

US20090027672A1 Axial Illumination for Capillary Electrophoresis 审中-公开
标题翻译：毛细管电泳的轴向照明
公开(公告)号：US20090027672A1
公开(公告)日：2009-01-29
申请号：US12240722
申请日：2008-09-29
申请人： Richard T. Reel , Eric S. Nordman
发明人： Richard T. Reel , Eric S. Nordman
IPC分类号： G01N21/59
CPC分类号： G01N21/64 , F21V5/00 , G01N21/645 , G01N27/44721 , G01N2021/6463 , G01N2201/068 , Y10T436/143333
摘要： System and method for fluorescent light excitation and detection from samples to enhance the numerical aperture and/or reduce the cross-talk of the fluorescent light.
摘要翻译：用于荧光灯激发和检测样品的系统和方法，以提高数值孔径和/或减少荧光灯的串扰。

9. 发明申请

US20080314431A1 Thermal cycler for PCR 有权
标题翻译：热循环仪用于PCR
公开(公告)号：US20080314431A1
公开(公告)日：2008-12-25
申请号：US12229404
申请日：2008-08-22
申请人： John G. Atwood , Adrian Fawcett , Keith S. Ferrara , Paul M. Hetherington , Richard W. Noreiks , Douglas E. Olsen , John R. Widomski , Charles M. Wittmer
发明人： John G. Atwood , Adrian Fawcett , Keith S. Ferrara , Paul M. Hetherington , Richard W. Noreiks , Douglas E. Olsen , John R. Widomski , Charles M. Wittmer
IPC分类号： H01L35/28
CPC分类号： B01L7/52 , B01L3/50853 , B01L2300/0829 , B01L2300/1822 , B01L2300/1827 , C12Q1/686 , F25B21/04 , F25B2321/021 , G01R27/205 , G05D23/1919 , H01L35/30 , Y10T137/1516
摘要： An instrument for performing highly accurate PCR employing an assembly, a heated cover, and an internal computer, is provided. The assembly is made up of a sample block, a number of Peltier thermal electric devices, and a heat sink, clamped together. A control algorithm manipulates the current supplied to thermoelectric coolers such that the dynamic thermal performance of a block can be controlled so that pre-defined thermal profiles of sample temperature can be executed. The sample temperature is calculated instead of measured using a design specific model and equations. The control software includes calibration diagnostics which permit variation in the performance of thermoelectric coolers from instrument to instrument to be compensated for such that all instruments perform identically. The block/heat sink assembly can be changed to another of the same or different design. The assembly carries the necessary information required to characterize its own performance in an on-board memory device, allowing the assembly to be interchangeable among instruments while retaining its precision operating characteristics.
摘要翻译：提供了一种使用组件，加热盖和内部计算机执行高精度PCR的仪器。组件由样品块，多个珀耳帖热电器件和散热器组成，夹在一起。控制算法控制提供给热电冷却器的电流，使得可以控制块的动态热性能，从而可以执行样本温度的预定义热分布。计算样品温度，而不是使用设计特定的模型和方程来测量。控制软件包括校准诊断，其允许从仪器到仪器的热电冷却器的性能的变化被补偿，使得所有仪器执行相同的操作。块/散热器组件可以改变为相同或不同设计的另一个。该组件携带必要的信息，以在车载存储设备中表征自己的性能，允许组件在仪器之间可互换，同时保持其精确的操作特性。

10. 发明授权

US07452510B2 Manually-operable multi-well microfiltration apparatus and method 有权
标题翻译：手动操作的多孔微滤装置及方法
公开(公告)号：US07452510B2
公开(公告)日：2008-11-18
申请号：US11338254
申请日：2006-01-24
申请人： Todd A. Weinfield , Gary Lim , Donald R. Sandell , Kevin S. Bodner , Mark Borodkin , Mark Oldham , Jon Hoshizaki
发明人： Todd A. Weinfield , Gary Lim , Donald R. Sandell , Kevin S. Bodner , Mark Borodkin , Mark Oldham , Jon Hoshizaki
IPC分类号： B01L11/00 , B01L3/00 , B01D63/00 , B65B43/42
CPC分类号： B01L3/50255 , B01D61/18 , B01J2219/00288 , B01J2219/00313 , B01J2219/00319 , B01J2219/00335 , B01J2219/00416 , B01J2219/00418 , B01J2219/00423 , B01L7/52 , B01L2200/025 , B01L2200/0631 , B01L2200/0689 , B01L2200/141 , B01L2300/0829 , B01L2300/1805 , B01L2400/049 , G01N1/4077 , G01N35/0099 , G01N35/028 , G01N35/1074 , G01N2035/00485 , G01N2035/102 , Y10T436/25375
摘要： A multi-well microfiltration apparatus and method are provided and feature a manual touch-off system for transferring pendent drops hanging from discharge-conduits of a discharge-conduit array to respective receiving wells or receiving holes of a corresponding receiving array, with minimum or no cross-contamination between the discharge conduits, or the receiving wells or receiving holes. The manual touch-off is achieved by manually shifting a carriage that supports one of the arrays, into a position whereat pendent drops of fluid hanging from the distal ends of the discharge conduits contact the inner sidewalls of the corresponding receiving wells or receiving holes of the receiving array.
摘要翻译：提供了一种多孔微滤装置和方法，其特征在于一种手动触发系统，用于将从排放管阵列的排放管道悬挂的悬挂液滴转移到相应接收孔的相应接收孔或接收孔的最小或不排放管道或接收井或接收孔之间的交叉污染。手动触发是通过将支撑其中一个阵列的托架手动地移动到从排出管道的远端悬挂的悬挂液滴与相应的接收井的内侧壁接触的位置或接收孔的接收孔接收阵列。

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