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公开(公告)号：US20080227124A1
公开(公告)日：2008-09-18
申请号：US12076235
申请日：2008-03-14
CPC分类号： C12Q1/6823 , B01J2219/00659 , B01J2219/00702 , B01J2219/00722 , B01J2219/00725 , C40B40/06 , C40B40/10 , C40B50/14 , G01N33/54393 , Y10S436/805 , Y10S436/823
摘要： Solid supports for chemiluminescent assays are provided. The solid support includes a plurality of probes covalently or physically attached to the support surface and a chemiluminescent enhancing moiety incorporated onto the surface or into the bulk of the support. The solid support can be a multi-layered support including an upper probe binding layer (e.g., an azlactone polymer layer or porous functional polyamide layer) adjacent to a cationic microgel layer. The azlactone-functional polymer can be a copolymer of dimethylacrylamide and vinylazlactone crosslinked with ethylenediamine. The cationic microgel layer can be a cross-linked quaternary onium salt containing polymer. A method and a kit for conducting chemiluminescent assays using the solid supports is also provided. The kit comprises a dioxetane substrate, a biopolymer probe-enzyme complex, and a solid support. The solid support can be an azlactone functional polymer layer adjacent to a cationic microgel layer; a porous polyamide functional layer adjacent to a cationic microgel layer; or a quaternized azlactone functional polymer layer.
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公开(公告)号：US07368296B2
公开(公告)日：2008-05-06
申请号：US10046730
申请日：2002-01-17
IPC分类号： G01N33/545 , G01N33/533 , C07C211/62
CPC分类号： C12Q1/6823 , B01J2219/00659 , B01J2219/00702 , B01J2219/00722 , B01J2219/00725 , C40B40/06 , C40B40/10 , C40B50/14 , G01N33/54393 , Y10S436/805 , Y10S436/823
摘要： Solid supports for chemiluminescent assays are provided. The solid support includes a plurality of probes covalently or physically attached to the support surface and a chemiluminescent enhancing moiety incorporated onto the surface or into the bulk of the support. The solid support can be a multi-layered support including an upper probe binding layer (e.g., an azlactone polymer layer or porous functional polyamide layer) adjacent to a cationic microgel layer. The azlactone-functional polymer can be a copolymer of dimethylacrylamide and vinylazlactone crosslinked with ethylenediamine. The cationic microgel layer can be a cross-linked quaternary onium salt containing polymer. A method and a kit for conducting chemiluminescent assays using the solid supports is also provided. The kit comprises a dioxetane substrate, a biopolymer probe-enzyme complex, and a solid support. The solid support can be an azlactone functional polymer layer adjacent to a cationic microgel layer; a porous polyamide functional layer adjacent to a cationic microgel layer; or a quaternized azlactone functional polymer layer.
摘要翻译： 提供了化学发光测定的固体支持物。 固体支持物包括共价或物理连接到支持物表面上的多个探针和结合到表面或载体的大部分中的化学发光增强部分。 固体支持物可以是包含邻近阳离子微凝胶层的上探针结合层（例如，吖内酯聚合物层或多孔功能性聚酰胺层）的多层载体。 吖内酯官能聚合物可以是与乙二胺交联的二甲基丙烯酰胺和乙烯基内酯的共聚物。 阳离子微凝胶层可以是交联的含季铵盐的聚合物。 还提供了使用固体支持物进行化学发光测定的方法和试剂盒。 试剂盒包括二氧杂环丁烷底物，生物聚合物探针 - 酶复合物和固体支持物。 固体支持物可以是与阳离子微凝胶层相邻的吖内酯官能聚合物层; 与阳离子微凝胶层相邻的多孔聚酰胺官能层; 或季铵化的吖内酯官能聚合物层。
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公开(公告)号：US07658288B2
公开(公告)日：2010-02-09
申请号：US11270325
申请日：2005-11-08
申请人： Yafei Liu , Haley O. Fiske , Steven M. Menchen
发明人： Yafei Liu , Haley O. Fiske , Steven M. Menchen
摘要： Disclosed, among other things, are packaged bisulfite solutions comprising bisulfite reagent in an oxygen-impermeable container and methods.
摘要翻译： 公开了包括在不透氧容器中的包含亚硫酸氢盐试剂的包装的亚硫酸氢盐溶液和方法。
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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”。
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公开(公告)号：US07364855B2
公开(公告)日：2008-04-29
申请号：US11119985
申请日：2005-05-02
CPC分类号： C12Q1/68 , C12Q1/6827 , C12Q1/6862 , C12Q2537/164 , C12Q2521/501
摘要： Ligation-based methods and kits are disclosed for determining the degree of methylation of one or more target nucleotides. In certain embodiments, the methylation status of one or more target nucleotides is determined by generating misligation products. In certain embodiments, at least one target nucleotide is amplified prior to the ligation reaction. In certain embodiments, at least one ligation product, at least one ligation product surrogate, at least one misligation product, at least one misligation product surrogate, or combinations thereof are amplified. In certain embodiments, one or more ligation probes comprise at least one nucleotide analog, at least one Modification, at least one mismatched nucleotide, or combinations thereof.
摘要翻译： 公开了基于连接的方法和试剂盒以确定一个或多个靶核苷酸的甲基化程度。 在某些实施方案中，通过产生错误产物来确定一个或多个靶核苷酸的甲基化状态。 在某些实施方案中，在连接反应之前扩增至少一种靶核苷酸。 在某些实施方案中，扩增至少一种连接产物，至少一种连接产物替代物，至少一种错误产物，至少一种错误产物替代物或其组合。 在某些实施方案中，一个或多个连接探针包含至少一个核苷酸类似物，至少一个修饰，至少一个错配核苷酸或其组合。
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公开(公告)号：US06504024B2
公开(公告)日：2003-01-07
申请号：US09881898
申请日：2001-06-14
IPC分类号： C07H1900
摘要： Propargylethoxyamino nucleosides are disclosed having the structure wherein R1 and R2 are —H, lower alkyl, or label; B is a 7-deazapurine, purine, or pyrimidine nucleoside base; W1 is —H or —OH; W2 is —OH or a moiety which renders the nucleoside incapable of forming a phosphodiester bond at the 3′-position; and W3 is —PO4, —P2O7, —P3O10, phosphate analog, or —OH. Additionally, a primer extension method is provided employing the above propargylethoxyamino nucleosides.
摘要翻译： 公开了具有其中R1和R2为-H，低级烷基或标记的结构的丙炔基乙氧基氨基核苷; B是7-脱氮嘌呤，嘌呤或嘧啶核苷碱基; W1是-H或-OH; W2是-OH或使核苷不能在3'-位形成磷酸二酯键的部分; W3是-PO 4，-P 2 O 7，-P 3 O 10，磷酸酯类似物或-OH。 另外，使用上述炔丙氧基氨基核苷提供引物延伸方法。
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公开(公告)号：US06358385B1
公开(公告)日：2002-03-19
申请号：US09228991
申请日：1999-01-11
IPC分类号： B01D5702
CPC分类号： G01N27/44752 , G01N27/44747
摘要： The invention provides uncharged water-soluble silica-adsorbing polymers for suppressing electroendoosmotic flow and to reduce analyte-wall interactions in capillary electrophoresis. In one aspect of the invention, one or more of such polymers are employed as components of a separation medium for the separation of biomolecules, such as polynucleotides, polysaccharides, proteins, and the like, by capillary electrophoresis. Generally, such polymers are characterized by (i) water solubility over the temperature range between about 20° C. to about 50° C., (ii) concentration in a separation medium in the range between about 0.001% to about 10% (weight/volume), (iii) molecular weight in the range of about 5×103 to about 1×106 daltons, and (iv) absence of charged groups in an aqueous medium having pH in the range of about 6 to about 9. In one embodiment, polymers of the invention are selected from the group consisting of polylactams, such as polyvinylpyrrolidone; N,N-disubstituted polyacrylamides; and N-substituted polyacrylamides. In accordance with the method of the invention, a sufficient amount of polymer adsorbs to the capillary surface to establish a zone of high viscosity that shields the analyte from the wall and impedes the movement of an electrical double layer under an electric field.
摘要翻译： 本发明提供了不带电的水溶性二氧化硅吸附聚合物，用于抑制毛细管电泳中的电渗流和降低毛细管电泳中的分析物 - 壁相互作用。 在本发明的一个方面，一种或多种这样的聚合物用作用于通过毛细管电泳分离生物分子例如多核苷酸，多糖，蛋白质等的分离介质的组分。 通常，这种聚合物的特征在于（i）在约20℃至约50℃之间的温度范围内的水溶性，（ii）分离介质中的浓度在约0.001％至约10％（重量） /体积），（iii）约5×10 3至约1×10 6道尔顿范围内的分子量，和（iv）在pH为约6至约9范围内的水性介质中不存在带电基团。在一个实施方案中， 本发明选自聚内酰胺类，如聚乙烯吡咯烷酮; N，N-二取代的聚丙烯酰胺; 和N-取代的聚丙烯酰胺。 根据本发明的方法，足够量的聚合物吸附到毛细管表面以建立一个高粘度的区域，从而将分析物与壁隔离并阻止双电层在电场下移动。
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公开(公告)号：US06248884B1
公开(公告)日：2001-06-19
申请号：US09325243
申请日：1999-06-03
IPC分类号： C07D31194
CPC分类号： C12Q1/6869 , C07D209/60 , C07D221/08 , C07D221/10 , C07D471/06 , C07D487/06 , C07D491/14 , C07H19/04 , C07H21/00 , C09B11/24 , C12Q1/68 , C12Q1/6816 , Y10S435/968 , Y10S435/975 , Y10S436/80 , Y10T436/143333
摘要： Extended rhodamine compounds exhibiting favorable fluorescence characteristics having the structure are disclosed. In addition, novel intermediates for synthesis of these dyes are disclosed, such intermediates having the structure In addition, methods of making and using the dyes as fluorescent labels are disclosed.
摘要翻译： 显示出具有有利结构的良好荧光特性的延伸罗丹明化合物。 此外，公开了用于合成这些染料的新型中间体，具有该结构的这种中间体。此外，公开了制备和使用染料作为荧光标记的方法。
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公开(公告)号：US06248568B1
公开(公告)日：2001-06-19
申请号：US09583068
申请日：1998-06-18
IPC分类号： C12P1934
摘要： Propargylethoxyamino nucleosides are disclosed having the structure wherein R1 and R2 are —H, lower alkyl, or label; B is a 7-deazapurine, purine, or pyrimidine nucleoside base; W1 is —H or —OH; W2 is —OH or a moiety which renders the nucleoside incapable of forming a phosphodiester bond at the 3′-position; and W3 is —PO4, —P2O7, —P3O10, phosphate analog, or —OH. Additionaly, a primer extension method is provided employing the above propargylethoxyamino nucleosides.
摘要翻译： 公开了具有其中R1和R2为-H，低级烷基或标记的结构的丙炔基乙氧基氨基核苷; B是7-脱氮嘌呤，嘌呤或嘧啶核苷碱基; W1是-H或-OH; W2是-OH或使核苷不能在3'-位形成磷酸二酯键的部分; W3是-PO 4，-P 2 O 7，-P 3 O 10，磷酸酯类似物或-OH。 另外，使用上述炔丙氧基氨基核苷提供引物延伸方法。
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公开(公告)号：US06197555B1
公开(公告)日：2001-03-06
申请号：US09461510
申请日：1999-12-14
IPC分类号： C12P1934
CPC分类号： C07H19/06 , C07H19/04 , C07H19/10 , C07H19/14 , C07H19/16 , C07H19/20 , C07H21/00 , C12Q1/6869 , C12Q2535/101
摘要： A nucleoside/tide compound having the structure NUC-L-S-LB/LG is described wherein NUC is a nucleoside/tide having a nucleobase portion B, L is a rigid linkage, S is a spacer; and LB/LG is a member of a linkage pair or a label. NUC is attached to L through B such that when B is a purine, L is attached to the 8-position of the purine, when B is 7-deazapurine, L is attached to the 7-position of the 7-deazapurine, and when B is pyrimidine, L is attached to the 5-position of the pyrimidine. In an important aspect of the invention, L has the structure wherein each of n, o and p are integers ranging from 0 to 3, and the sum of n, o and p is at least 2, and each of W, X, Y and Z is selected from the group consisting of carbon and nitrogen. The invention further includes polynucleotide compounds comprising the nucleoside/tide, and primer extension methods utilizing the nucleoside/tide, particularly when used in combination with certain mutant polymerase enzymes.
摘要翻译： 具有所述结构的核苷/潮汐化合物，其中NUC是具有核碱基部分B的核苷/ /，L是刚性键，S是间隔基; 而LB / LG是连锁对或标签的成员。 N与B连接，B为嘌呤时，L为嘌呤8位时，B为7-脱氮嘌呤时，L为7-脱氮嘌呤的7-位， B是嘧啶，L连接到嘧啶的5位上。 在本发明的一个重要方面，L具有其中n，o和p各自为0至3的整数，n，o和p之和至少为2，W，X，Y和 Z选自碳和氮。 本发明还包括包含核苷/潮汐的多核苷酸化合物，以及利用核苷/潮汐的引物延伸方法，特别是当与某些突变型聚合酶组合使用时。
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