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1. 发明授权

US5514543A Method and probe composition for detecting multiple sequences in a single assay 失效
标题翻译：用于在单个测定中检测多个序列的方法和探针组合物
公开(公告)号：US5514543A
公开(公告)日：1996-05-07
申请号：US102372
申请日：1993-08-04
申请人： Paul D. Grossman , Steven Fung , Steven M. Menchen , Sam L. Woo , Emily S. Win-Deen
发明人： Paul D. Grossman , Steven Fung , Steven M. Menchen , Sam L. Woo , Emily S. Win-Deen
IPC分类号： C12N15/09 , B01D57/02 , C12Q1/68 , G01N27/447 , G01N30/88 , G01N33/566
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 one embodiment of the invention, a plurality of different-sequence probe pairs are added to a target polynucleotide, where each probe pair includes two polynucleotide probe elements which are complementary in sequence to adjacent portions of a selected one of the target sequences in the target polynucleotide. In each probe pair, one of the probe elements contains a non-polynucleotide polymer chain which imparts a distinctive mobility to the associated probe pair, when the elements in the pair are ligated. The other element in the pair contains a detectable reporter label. After the probe pairs have been allowed to hybridize with the target polynucleotide, the hybridized polynucleotides are treated under conditions effective to ligate the end subunits of target-bound probe elements when their end subunits are base-paired with adjacent target bases. The ligated probe pairs are then released from the target polynucleotide and separated electrophoretically in a sieving matrix, or chromatographically.
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。在本发明的一个实施方案中，将多个不同序列探针对加入到靶多核苷酸中，其中每个探针对包含两个多核苷酸探针元件，该多核苷酸探针元件与目标中所选择的一个目标序列的相邻部分互补多核苷酸。在每个探针对中，探针元件中的一个包含非多核苷酸聚合物链，当对中的元件被连接时，赋予相关探针对的特异性移动性。该对中的另一个元素包含可检测的报告物标记。在探针对已被允许与靶多核苷酸杂交后，杂交的多核苷酸在有效连接靶结合探针元件的末端亚基的条件下处理，当它们的末端亚基与相邻的靶基碱基配对时。然后将连接的探针对从目标多核苷酸中释放，并在筛分基质中电泳分离，或进行色谱分离。

2. 发明授权

US5624800A Method of DNA sequencing employing a mixed DNA-polymer chain probe 失效
标题翻译：使用混合DNA-聚合物链探针进行DNA测序的方法
公开(公告)号：US5624800A
公开(公告)日：1997-04-29
申请号：US447174
申请日：1995-05-19
申请人： Paul D. Grossman , Steven Fung , Steven M. Menchen , Sam L. Woo , Emily S. Winn-Deen
发明人： Paul D. Grossman , Steven Fung , Steven M. Menchen , Sam L. Woo , Emily S. Winn-Deen
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。

3. 发明授权

US5580732A Method of DNA sequencing employing a mixed DNA-polymer chain probe 失效
标题翻译：使用混合DNA-聚合物链探针进行DNA测序的方法
公开(公告)号：US5580732A
公开(公告)日：1996-12-03
申请号：US296880
申请日：1994-08-26
申请人： Paul D. Grossman , Steven Fung , Steven M. Menchen , Sam L. Woo , Emily S. Winn-Deen
发明人： Paul D. Grossman , Steven Fung , Steven M. Menchen , Sam L. Woo , Emily S. Winn-Deen
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-sieiving 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进行分级，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。

4. 发明授权

US5470705A Probe composition containing a binding domain and polymer chain and methods of use 失效
标题翻译：含有结合域和聚合物链的探针组合物及其使用方法
公开(公告)号：US5470705A
公开(公告)日：1995-11-28
申请号：US866018
申请日：1992-04-07
申请人： Paul D. Grossman , Steven Fung , Steven M. Menchen , Sam L. Woo , Emily S. Winn-Deen
发明人： Paul D. Grossman , Steven Fung , Steven M. Menchen , Sam L. Woo , Emily S. Winn-Deen
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6816 , C12Q1/6825 , C12Q1/6827 , C12Q1/686 , C12Q1/6862 , C12Q2600/156 , G01N27/447 , 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。

5. 发明授权

US6096723A 4,7-dichlorofluorescein dyes as molecular probes 有权
标题翻译： 4,7-二氯荧光素染料作为分子探针
公开(公告)号：US6096723A
公开(公告)日：2000-08-01
申请号：US273655
申请日：1999-03-23
申请人： Steven M. Menchen , Linda G. Lee , Charles R. Connell , N. Davis Hershey , Vergine Chakerian , Sam L. Woo , Steven Fung
发明人： Steven M. Menchen , Linda G. Lee , Charles R. Connell , N. Davis Hershey , Vergine Chakerian , Sam L. Woo , Steven Fung
IPC分类号： C07D311/82 , C07H21/00 , C09B11/08 , C12Q1/68 , A01N43/04
CPC分类号： C07H21/00 , C07D311/82 , C09B11/08 , C12Q1/6816 , C12Q1/6869 , A61K48/00 , Y10S435/968 , Y10S436/80 , Y10T436/143333 , Y10T436/145555 , Y10T436/147777
摘要： Long wavelength, narrow emission bandwidth fluorecein dyes are provided for detecting spacially overlapping target substances. The dyes comprise 4,7-dichlorofluoresceins, and particularly 2',4',5', 7'-tetrachloro-4,7-dichloro-5- (and 6-) carboxyfluoresceins. Methods and kits for using the dyes in DNA analysis are provided.
摘要翻译：提供长波长，窄发射带宽荧光素染料用于检测空间重叠的目标物质。染料包括4,7-二氯荧光素，特别是2'，4'，5'，7'-四氯-4,7-二氯-5-（和6-）羧基荧光素。提供了在DNA分析中使用染料的方法和试剂盒。

6. 发明授权

US5885778A 4, 7-dichlorofluorescein dyes as molecular probes 失效
标题翻译： 4,7-二氯荧光素染料作为分子探针
公开(公告)号：US5885778A
公开(公告)日：1999-03-23
申请号：US905855
申请日：1997-08-04
申请人： Steven M. Menchen , Linda G. Lee , Charles R. Connell , N. Davis Hershey , Vergine Chakerian , Sam L. Woo , Steven Fung
发明人： Steven M. Menchen , Linda G. Lee , Charles R. Connell , N. Davis Hershey , Vergine Chakerian , Sam L. Woo , Steven Fung
IPC分类号： C07D311/82 , C07H21/00 , C09B11/08 , C12Q1/68 , C07D311/78 , C07D311/94
CPC分类号： C07H21/00 , C07D311/82 , C09B11/08 , C12Q1/6816 , C12Q1/6869 , A61K48/00 , Y10S435/968 , Y10S436/80 , Y10T436/143333 , Y10T436/145555 , Y10T436/147777
摘要： Long wavelength, narrow emission bandwidth fluorecein dyes are provided for detecting spacially overlapping target substances. The dyes comprise 4,7-dichlorofluoresceins, and particularly 2',4',5',7'-tetrachloro-4,7-dichloro-5- (and 6-) carboxyfluoresceins. Methods and kits for using the dyes in DNA analysis are provided.
摘要翻译：提供长波长，窄发射带宽荧光素染料用于检测空间重叠的目标物质。染料包括4,7-二氯荧光素，特别是2'，4'，5'，7'-四氯-4,7-二氯-5-（和6-）羧基荧光素。提供了在DNA分析中使用染料的方法和试剂盒。

7. 发明授权

US4965349A Method of synthesizing oligonucleotides labeled with ammonia-labile groups on solid phase supports 失效
标题翻译：在固相载体上合成用氨不稳定基团标记的寡核苷酸的方法
公开(公告)号：US4965349A
公开(公告)日：1990-10-23
申请号：US138287
申请日：1987-12-24
申请人： Sam L. Woo , Steven M. Menchen , Steven Fung
发明人： Sam L. Woo , Steven M. Menchen , Steven Fung
IPC分类号： C12N15/09 , C07F9/24 , C07H21/00
CPC分类号： C07H21/00 , C07F9/24
摘要： The invention provides a novel cleavage reagent for hydrolysing base-labile linking groups between a solid phase support and oligonucleotides. The cleavage reagent comprises a lower alkyl alcohol, water, and a non-nuccleophilic hindered alkylamine containing from 3 to 6 carbon atoms in a ratio of about 1:1:1 to about 1:3:1, respectively. An important property of the cleavage reagent is that it preserves the fluorescent characteristics of rhodamine dyes during cleavage, thereby making it possible to completely synthesize rhodamine-labeled oligonucleotides by solid phase methods. Rhodamine phosphoramidites are provided to further enhance the efficiency of synthesizing rhodamine-labeled oligonucleotides by such methods.

8. 发明授权

US5366860A Spectrally resolvable rhodamine dyes for nucleic acid sequence determination 失效
标题翻译：用于核酸序列测定的光谱可分辨的罗丹明染料
公开(公告)号：US5366860A
公开(公告)日：1994-11-22
申请号：US415050
申请日：1989-09-29
申请人： B. John Bergot , Vergine Chakerian , Charles R. Connell , J. Scott Eadie , Steven Fung , N. Davis Hershey , Linda G. Lee , Steven M. Menchen , Sam L. Woo
发明人： B. John Bergot , Vergine Chakerian , Charles R. Connell , J. Scott Eadie , Steven Fung , N. Davis Hershey , Linda G. Lee , Steven M. Menchen , Sam L. Woo
IPC分类号： C09B69/10 , C07H19/10 , C07H19/20 , C07H21/00 , C09B11/24 , C12Q1/68
CPC分类号： C12Q1/6869 , C07H21/00 , C09B11/24
摘要： A spectrally resolvable set of rhodamine dyes are provided for use in the chain termination method of nucleic acid sequencing. A different rhodamine dye from the group consisting of tetramethylrhodamine, rhodamine X, rhodamine 6G, and rhodamine 110 is attached to the base of each of the dideoxynucleotides used in the sequencing method by way of an alkynylamino linker. Preferably, the labeled dideoxynucleotides are incorporated into the growing DNA chains by Taq DNA polymerase.
摘要翻译：提供光谱可分辨的罗丹明染料组合用于核酸测序的链终止方法。通过炔基氨基接头将与四氯乙酰胺，罗丹明X，罗丹明6G和罗丹明110组成的组中的不同的罗丹明染料连接到用于测序方法的每个双脱氧核苷酸的碱基上。优选地，标记的双脱氧核苷酸通过Taq DNA聚合酶掺入生长的DNA链中。

9. 发明授权

US4855225A Method of detecting electrophoretically separated oligonucleotides 失效
标题翻译：检测电泳分离的寡核苷酸的方法
公开(公告)号：US4855225A
公开(公告)日：1989-08-08
申请号：US827348
申请日：1986-02-07
申请人： Steven Fung , Sam L. Woo , Richard P. Haugland , Steven M. Menchen , Charles R. Connell
发明人： Steven Fung , Sam L. Woo , Richard P. Haugland , Steven M. Menchen , Charles R. Connell
IPC分类号： C07H21/04 , C07H1/06 , C07H21/00 , C12Q1/68 , G01N27/26 , G01N27/447 , G01N33/50 , G01N33/58
CPC分类号： G01N33/582 , C07H21/00 , C12Q1/6816 , C12Q1/6874
摘要： A method is provided for detecting up to four classes of oligonucleotides which have been separated by gel electrophoresis. The method entails labeling members of each class of oligonucleotide with dyes selected from separate sets of dyes so that members of the same class are labeled with dyes from the same set. The four sets of dyes of the invention consist of derivatives of fluorescein, 2',7'-dimethoxy-4', 5'-dichlorofluorescein, tetramethylrhodamine, and rhodamine X carboxylic or sulfonic acid, respectively. Dyes from these sets are spectrally resolvable under conditions of gel electrophoresis.
摘要翻译：提供了一种用于检测多达四类通过凝胶电泳分离的寡核苷酸的方法。该方法需要用选自单独的染料组的染料标记每一类寡核苷酸的成员，使得同一类的成员用来自同一组的染料标记。本发明的四组染料分别由荧光素，2'，7'-二甲氧基-4'，5'-二氯荧光素，四甲基罗丹明和罗丹明X羧酸或磺酸的衍生物组成。来自这些组的染料在凝胶电泳条件下是光谱可分辨的。

10. 发明授权

US5231191A Rhodamine phosphoramidite compounds 失效
标题翻译：罗丹明亚磷酰胺化合物
公开(公告)号：US5231191A
公开(公告)日：1993-07-27
申请号：US601961
申请日：1990-10-22
申请人： Sam L. Woo , Steven M. Menchen , Steven Fung
发明人： Sam L. Woo , Steven M. Menchen , Steven Fung
IPC分类号： C07F9/24 , C07H21/00
CPC分类号： C07H21/00 , C07F9/24
摘要： The invention provides a novel cleavage reagent for hydrolysing base-labile linking groups between a solid phase support and oligonucleotides. The cleavage reagent comprises a lower alkyl alcohol, water, and a non-nuccleophilic hindered alkylamine containing from 3 to 6 carbon atoms in a ratio of about 1:1:1 to about 1:3:1, respectively. An important property of the cleavage reagent is that it preserves the fluorescent characteristics of rhodamine dyes during cleavage, thereby making it possible to completely synthesize rhodamine-labeled oligonucleotides by solid phase methods. Rhodamine phosphoramidites are provided to further enhance the efficiency of synthesizing rhodamine-labeled oligonucleotides by such methods.
摘要翻译：本发明提供了一种用于水解固相载体和寡核苷酸之间的碱不稳定连接基团的新型裂解试剂。裂解试剂包含分别含有约1：1：1至1：3：1的3至6个碳原子的低级烷基醇，水和非亲核的受阻烷基胺。裂解试剂的重要性质是在切割期间保留罗丹明染料的荧光特性，从而可以通过固相方法完全合成罗丹明标记的寡核苷酸。提供罗丹明亚磷酰胺以通过这些方法进一步提高合成罗丹明标记的寡核苷酸的效率。

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