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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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2.发明申请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
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。
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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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10.发明授权
公开(公告)号：US5624800A
公开(公告)日：1997-04-29
申请号：US447174
申请日：1995-05-19
IPC分类号： C12N15/09 , B01D57/02 , C12Q1/68 , G01N27/447 , G01N30/88 , G01N33/566 , C12P19/34
CPC分类号： C12Q1/6869 , C12Q1/6816 , C12Q1/6827 , C12Q1/6858 , C12Q1/686 , C12Q1/6862 , C12Q1/6818 , Y10T436/143333
摘要： Method and composition for detecting one or more selected polynucleotide regions in a target polynucleotide. In the method, a mixture of sequence-specific probes are reacted with the target polynucleotide under hybridization conditions, and the hybridized probes are treated to selectively modify those probes which are bound to the target polynucleotide in a base-specific manner. The resulting labeled probes include a polymer chain which imparts to each different-sequence probe, a distinctive ratio of charge/translational frictional drag, and a detectable label. The labeled probes are fractionated by electrophoresis in a non-sieving matrix, and the presence of one or more selected sequences in the target polynucleotide are detected according to the observed electrophoretic migration rates of the labeled probes in a non-sieving medium.
摘要翻译： 用于检测靶多核苷酸中的一个或多个选择的多核苷酸区域的方法和组合物。 在该方法中，将序列特异性探针的混合物与靶多核苷酸在杂交条件下反应，并对杂交的探针进行处理以选择性地修饰以碱基特异性方式结合靶多核苷酸的那些探针。 所得到的标记探针包括赋予每个不同序列探针的聚合物链，电荷/平移摩擦阻力的独特比例和可检测标记。 标记的探针通过在非筛选基质中的电泳分级分离，并且根据在非筛选培养基中标记的探针的观察到的电泳迁移速率来检测靶多核苷酸中一个或多个选定序列的存在。
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