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11.发明授权
公开(公告)号：US5612199A
公开(公告)日：1997-03-18
申请号：US221662
申请日：1994-04-01
CPC分类号： C12Q1/686 , C12N15/10 , C12Q1/6844 , C12Q1/6853
摘要： A method is disclosed for extending an extender probe to produce a single stranded polydeoxynucleotide that is free of unreacted extender probe and has two segments that are non-contiguous and complementary with each other. The method comprises the steps of (1) providing in combination (a) a polynucleotide having two non-contiguous, non-complementary nucleotide sequences S1 and S2 wherein S2 is 5' of S1 and is at least ten deoxynucleotides long, (b) an extender probe comprised of two deoxynucleotide sequences, wherein the sequence at the 3'-end of the extender probe (EP1) is hybridizable with S1 and the other of the deoxynucleotide sequences (EP2) is substantially identical to S2 and (c) means for modifying the 3'-end of extender probe that does not hybridize with the polynucleotide and (2) extending the extender probe along the polynucleotide wherein extender probe not hybridized to the polynucleotide becomes modified at its 3'-end.
摘要翻译： 公开了一种用于延伸扩增器探针以产生不含未反应的扩增体探针的单链多脱氧核苷酸并具有彼此不连续和互补的两个区段的方法。 该方法包括以下步骤：（1）组合（a）具有两个非连续的，非互补的核苷酸序列S1和S2的多核苷酸，其中S2是S1的5'，并且长至少十个脱氧核苷酸，（b） 扩增物探针由两个脱氧核苷酸序列组成，其中扩增体探针（EP1）的3'末端的序列可与S1杂交，而另一个脱氧核苷酸序列（EP2）与S2基本相同，（c）修饰的手段 不与多核苷酸杂交的扩增体探针的3'末端和（2）沿多核苷酸延伸扩增体探针，其中不与多核苷酸杂交的扩增物探针在其3'末端被修饰。
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公开(公告)号：US5508178A
公开(公告)日：1996-04-16
申请号：US194140
申请日：1994-02-09
CPC分类号： C12Q1/6858 , Y10S435/81
摘要： A method is disclosed for determining the presence of a polynucleotide analyte in a sample suspected of containing the analyte. The method comprises (a) forming as a result of the presence of an analyte a single stranded polynucleotide comprising a target polynucleotide binding sequence flanked by first and second polynucleotide sequences that differ from the sequence of the analyte or a sequence complementary to the analyte sequence, (b) forming multiple copies of the single stranded polynucleotide, and (c) detecting the single stranded polynucleotide. Also disclosed is a method of producing at least one copy of a single stranded polynucleotide. The method comprises (a) forming in the presence of nucleoside triphosphates and template dependent polynucleotide polymerase an extension of a polynucleotide primer at least the 3'-end of which has at least a 10 base sequence hybridizable with a second sequence flanking the 3'-end of the single stranded polynucleotide, the second sequence being partially or fully complementary with at least a 10 base first sequence flanking the 5' end of the single stranded polynucleotide, (b) dissociating the extended polynucleotide primer and the single stranded polynucleotide, (c) repeating step a and (d) dissociating the extended polynucleotide primer and the copy of the single stranded polynucleotide.
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13.发明授权Method for preventing amplification of nucleic acid contaminants in amplification mixtures using nuclease-receptor conjugates 失效使用核酸酶 - 受体共轭物防止扩增混合物中核酸污染物扩增的方法
公开(公告)号：US5605796A
公开(公告)日：1997-02-25
申请号：US277547
申请日：1994-07-19
申请人： Yan Chen , Samuel J. Rose , Edwin F. Ullman
发明人： Yan Chen , Samuel J. Rose , Edwin F. Ullman
CPC分类号： C12Q1/6848
摘要： Methods and kits are disclosed for preventing amplification of contaminating copies of nucleic acids during in amplification of a nucleic acid suspected of being present in a sample. Modified nucleotides that render copies of the nucleic acid bindable by a member of a specific binding pair, such as a receptor, which does not bind to the nucleic acid, are incorporated into copies of the nucleic acid that are produced during the amplification. The sample is combined with an enzyme conjugate, usually a receptor bound to a nuclease, under conditions wherein prior to the amplification the member of a specific binding pair binds to the copies and the enzyme degrades the copies but not the nucleic acid. The methods and kits have particular application to the determination of a nucleic acid analyte.
摘要翻译： 公开了用于在怀疑存在于样品中的核酸扩增期间防止扩增核酸污染拷贝的方法和试剂盒。 通过特异性结合对的成员（例如不与核酸结合的受体）结合核酸的拷贝的修饰的核苷酸被并入在扩增过程中产生的核酸的拷贝中。 样品与酶缀合物（通常是与核酸酶结合的受体）在条件下结合，在扩增前，特异性结合对的成员与拷贝结合，并且酶降解拷贝而不是核酸。 所述方法和试剂盒特别适用于测定核酸分析物。
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公开(公告)号：US5914230A
公开(公告)日：1999-06-22
申请号：US771624
申请日：1996-12-20
申请人： Yen Ping Liu , Rajesh D. Patel , Nurith Kurn , Claire Lin , Samuel J. Rose , Edwin F. Ullman
发明人： Yen Ping Liu , Rajesh D. Patel , Nurith Kurn , Claire Lin , Samuel J. Rose , Edwin F. Ullman
IPC分类号： G01N33/53 , C12N15/09 , C12Q1/68 , G01N33/566 , C12P19/34
CPC分类号： C12Q1/6816 , C12Q1/6813 , C12Q1/6818 , C12Q1/686
摘要： The present invention relates to a method for detecting or amplifying and detecting a target polynucleotide sequence. The method comprises providing in combination (i) a medium suspected of containing the target polynucleotide sequence, (ii) all reagents required for conducting an amplification of the target polynucleotide sequence when amplification is desired, and (iii) two oligonucleotide probes capable of binding to a single strand of the product of the amplification. At least one of the probes has two sequences that either (i) are non-contiguous and bind to contiguous or non-contiguous sites on the single strand or (ii) can bind to non-contiguous sites on the single strand. Each probe may contain a label. The combination is subjected to conditions for amplifying the target polynucleotide sequence. Next, the combination is subjected to conditions under which both of the probes hybridize to one of the strands to form a termolecular complex, which is detected by means of the label.
摘要翻译： 本发明涉及检测或扩增和检测靶多核苷酸序列的方法。 所述方法包括组合（i）怀疑含有靶多核苷酸序列的培养基，（ii）当需要扩增时进行靶多核苷酸序列扩增所需的所有试剂，和（iii）能够结合到 单链的扩增产物。 至少一个探针具有以下两个序列：（i）不连续并且结合单链上的连续或非连续位点，或（ii）可以结合单链上的非连续位点。 每个探针可能包含一个标签。 将该组合进行扩增靶多核苷酸序列的条件。 接下来，将该组合进行两个探针与其中一条链杂交以形成通过标记检测的分子复合物的条件。
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公开(公告)号：US06294323B1
公开(公告)日：2001-09-25
申请号：US08046682
申请日：1993-04-14
申请人： Edwin F. Ullman , Samuel J. Rose
发明人： Edwin F. Ullman , Samuel J. Rose
IPC分类号： C07H2104
CPC分类号： C12Q1/6853 , C12Q1/6844 , C12Q2537/157 , C12Q2525/161 , C12Q2525/101
摘要： A method is disclosed for producing at least one copy of a pair of complementary single stranded polynucleotides. The method comprises forming, in the presence of nucleoside triphosphates and template dependent polynucleotide polymerase along each of the complementary single stranded polynucleotides, an extension of a polynucleotide primer. The polynucleotide primer is comprised of at least a sequence of 16 nucleotides terminating at its 3′ end in a 2 to 9 nucleotide sequence (S1), which is complementary with the 3′ ends of both of the complementary single stranded polynucleotides. The polynucleotide primer has at least an 8 nucleotide sequence (S2) that is 5′ of S1, where S2 is 50 to 80% complementary to the nucleotide sequences contiguous with the 3′ ends of the complementary single stranded polynucleotides. The extended polynucleotide primer and the single stranded polynucleotides are then dissociated.
摘要翻译： 公开了用于产生一对互补单链多核苷酸的至少一个拷贝的方法。 所述方法包括在存在核苷三磷酸和模板依赖多核苷酸聚合酶的情况下沿着每个互补单链多核苷酸形成多核苷酸引物的延伸。 多核苷酸引物由至少一个16个核苷酸的序列组成，其在3'末端以2至9个核苷酸序列（S1）终止，其与两个互补单链多核苷酸的3'末端互补。 多核苷酸引物具有至少8个S1的5'序列（S2），其中S2与互补单链多核苷酸的3'末端连接的核苷酸序列互补50至80％。 然后将延伸的多核苷酸引物和单链多核苷酸解离。
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公开(公告)号：US07635571B2
公开(公告)日：2009-12-22
申请号：US09732047
申请日：2000-12-07
IPC分类号： G01N33/00
CPC分类号： G01N33/542 , G01N33/531
摘要： Methods are disclosed for determining minute quantities of an analyte in a medium suspected of containing the analyte. One method comprises treating a medium suspected of containing an analyte under conditions such that the analyte, if present, causes a substrate having an oxidant cleavable linker and a photosensitizer to come into close proximity. The photosensitizer generates singlet oxygen which oxidatively cleaves the linker to form a product which can be detected in a sandwich detection assay such as LOCI. The amount of product detected is related to the amount of analyte in the medium. Compositions, kits, and compounds are also disclosed.
摘要翻译： 公开了用于确定怀疑含有分析物的介质中微量分析物的方法。 一种方法包括处理怀疑含有分析物的介质，使得分析物（如果存在的话）引起具有氧化剂可切割接头和光敏剂的底物接近。 光敏剂产生单线态氧，其氧化地切割接头以形成可在三明治检测测定（例如LOCI）中检测的产物。 所检测的产物量与培养基中分析物的量有关。 还公开了组合物，试剂盒和化合物。
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公开(公告)号：US07022529B2
公开(公告)日：2006-04-04
申请号：US10745972
申请日：2003-12-24
IPC分类号： G01N33/543
CPC分类号： G01N33/587 , C12Q1/6816 , Y10T436/10 , C12Q2563/155 , C12Q2523/319
摘要： Methods, compositions and kits are disclosed. The compositions are light emitting and comprise a polymeric matrix having dissolved therein a photoactive compound. The composition has the characteristic that, after activation of the photoactive compound, the rate of decrease in the intensity of light emission at any time during a 20-fold decrease in the intensity is proportional to the intensity of the light emission. In one embodiment the polymeric matrix is comprised of particles of about 20 nm to about 100 μm in diameter to which is bound a specific binding pair member. The particles generally comprise a polymeric matrix having dissolved therein about 1 to about 20% by weight of a dopant. The compositions may be used in methods for determining an analyte. A combination is provided comprising (1) a medium suspected of containing the analyte, (2) and the aforementioned composition. The photoactive substance is activated and the effect of the activating on the optical properties of the combination is detected. The presence and amount of the effect is related to the presence and amount of the analyte in the medium. Also disclosed are kits for use in an assay.
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公开(公告)号：US06783947B1
公开(公告)日：2004-08-31
申请号：US09393579
申请日：1999-09-09
IPC分类号： G01N33546
CPC分类号： G01N33/6842 , C07F9/113 , C07H15/10 , C12Q1/42 , G01N33/6815 , Y02P20/55 , Y10S435/975 , Y10S436/815 , Y10S436/825
摘要： Alpha-haloketones are useful alkylating agents for coupling to sulfhydryl-containing biomolecules. However, they react spontaneously with water, alkali and organic bases and therefore cannot be stored for extended periods of time in aqueous solutions, particularly in the presence of proteins at physiological pH. The present invention provides novel solutions to these problems, however, as novel compounds and compositions comprising protected haloketones are disclosed herein. Methods of preparing and using protected haloketones which are useful in a variety of applications—e.g., in assays and conjugation reactions—are also disclosed herein.
摘要翻译： α-卤代酮是用于与含巯基的生物分子偶联的有用的烷基化剂。 然而，它们与水，碱和有机碱发生自发反应，因此不能在水溶液中储存较长时间，特别是在生理pH值的蛋白质存在下。 然而，本发明提供了这些问题的新颖解决方案，因为本文公开了包含受保护的卤代酮的新型化合物和组合物。 制备和使用可用于各种应用（例如测定和缀合反应）的保护的卤代酮的方法也在本文中公开。
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公开(公告)号：US06627406B1
公开(公告)日：2003-09-30
申请号：US09568786
申请日：2000-05-10
申请人： Sharat Singh , Vivian Xiao , Ian Gibbons , Travis Boone , Torleif Ove Bjornson , Herbert H. Hooper , Edwin F. Ullman
发明人： Sharat Singh , Vivian Xiao , Ian Gibbons , Travis Boone , Torleif Ove Bjornson , Herbert H. Hooper , Edwin F. Ullman
IPC分类号： G01N3353
CPC分类号： B01L3/5027 , B01F13/0059 , B01J19/16 , B01J2219/00317 , B01L3/5025 , B01L3/502723 , B01L2200/0642 , B01L2200/142 , B01L2300/0861 , B01L2400/0406 , B01L2400/0415 , B01L2400/0688 , C40B60/14 , G01N1/34 , G01N2035/00237 , G01N2035/00287
摘要： Devices and methods are provided using microfluidic devices for manipulating small volumes and determining a variety of chemical and physical events. The devices rely upon an opening to the atmosphere of a small volume in a zone, where a sample is placed in the zone where evaporation can occur. The zone is maintained in contact with a liquid medium that serves to replenish the liquid in the zone and maintain the composition of the mixture in the zone substantially constant. The diffusion of components in the zone is restricted during the course of the determination by the liquid flux into the zone.
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公开(公告)号：US06555389B1
公开(公告)日：2003-04-29
申请号：US09470677
申请日：1999-12-23
申请人： Edwin F. Ullman , Sharat Singh , Ian Gibbons , Travis Boone , Torlief Bjornson
发明人： Edwin F. Ullman , Sharat Singh , Ian Gibbons , Travis Boone , Torlief Bjornson
IPC分类号： G01N33558
CPC分类号： B01L3/5027 , B01F13/0059 , B01J19/16 , B01J2219/00317 , B01L3/5025 , B01L3/502723 , B01L2200/0642 , B01L2200/142 , B01L2300/0861 , B01L2400/0406 , B01L2400/0415 , B01L2400/0688 , C40B60/14 , G01N1/34 , G01N2035/00237 , G01N2035/00287
摘要： Devices and methods are provided using microfluidic devices for manipulating small volumes and determining a variety of chemical and physical events. The devices rely upon an opening to the atmosphere of a small volume in a zone, where a sample is placed in the zone where evaporation can occur. The zone is maintained in contact with a liquid medium that serves to replenish the liquid in the zone and maintain the composition of the mixture in the zone substantially constant. The diffusion of components in the zone is restricted during the course of the determination by the liquid flux into the zone.
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