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1. 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。

2. 发明授权

US06649598B2 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：US06649598B2
公开(公告)日：2003-11-18
申请号：US09949444
申请日：2001-09-07
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： A01N4304
CPC分类号： C07H21/00 , A61K48/00 , C07D311/82 , C09B11/08 , C12Q1/6816 , C12Q1/6869 , Y10S435/968 , Y10S436/80 , Y10T436/143333 , Y10T436/145555 , Y10T436/147777 , C12Q2563/107
摘要： Long wavelength, narrow emission bandwidth fluorecein dyes are provided for detecting specially 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，用于检测特别重叠的目标物质。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。

3. 发明授权

US06403812B1 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：US06403812B1
公开(公告)日：2002-06-11
申请号：US09580754
申请日：2000-05-30
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： C07D31182
CPC分类号： C07H21/00 , A61K48/00 , C07D311/82 , C09B11/08 , C12Q1/6816 , C12Q1/6869 , Y10S435/968 , Y10S436/80 , Y10T436/143333 , Y10T436/145555 , Y10T436/147777 , C12Q2563/107
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-dicloro-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。

4. 发明授权

US5654442A 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：US5654442A
公开(公告)日：1997-08-05
申请号：US400780
申请日：1995-03-08
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
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。

5. 发明授权

US5212304A Amino-derivatized phosphoramidite linking agents 失效
标题翻译：氨基衍生的亚磷酰胺连接剂
公开(公告)号：US5212304A
公开(公告)日：1993-05-18
申请号：US216768
申请日：1988-07-08
申请人： Steven Fung , Sam L. Woo , Lloyd M. Smith
发明人： Steven Fung , Sam L. Woo , Lloyd M. Smith
IPC分类号： C07H1/00 , C07F9/09 , C07F9/24 , C07F9/572 , C07F9/59 , C07F9/6509 , C07F9/6584 , C07H19/10 , C07H19/20 , C07H21/00 , C08B37/00 , C08B37/02
CPC分类号： C07F9/2416 , A61K47/48061 , A61K47/48084 , C07F9/2408 , C07F9/65842 , C07H21/00 , C08B37/0021 , C08B37/0039
摘要： The compounds of the invention include novel linking agents comprising 2-substituted-3-protected-1,3,2-oxazaphosphacycloalkanes and their phosphoramidite precursors. The compounds of the invention further include conjugates of the above linking agents with oligonucleotides and polymer supports. The compounds of the present invention are useful for linking organic moieties, such as fluorescent or chromogenic dyes, to polymer supports and oligonucleotides, particularly single- and double-stranded DNA and RNA fragments.
摘要翻译：本发明的化合物包括包含2-取代-3-保护的1,3,2-氧杂氮杂环烷烃及其亚磷酰胺前体的新型交联剂。本发明的化合物还包括上述连接剂与寡核苷酸和聚合物载体的缀合物。本发明的化合物可用于将有机部分如荧光或显色染料与聚合物载体和寡核苷酸，特别是单链和双链DNA和RNA片段连接。

6. 发明授权

US5625052A Method for preparing 3'-nitrogen containing polynucleotides 失效
标题翻译：制备含3'-氮的多核苷酸的方法
公开(公告)号：US5625052A
公开(公告)日：1997-04-29
申请号：US576206
申请日：1995-12-21
申请人： Sam L. Woo , Steven Fung
发明人： Sam L. Woo , Steven Fung
IPC分类号： C12N15/09 , C07C217/20 , C07C271/10 , C07C271/16 , C07C271/66 , C07C317/18 , C07C333/04 , C07H21/00 , C07H21/04 , C07H1/00
CPC分类号： C07H21/00 , Y02P20/55
摘要： A method for synthesizing a polynucleotide having a nitrogen atom at the 3'-end is disclosed. The methods include the following steps. A solid support reagent is provided having the structure shown below: ##STR1## where T is an acid-cleavable hydroxyl protecting group, e.g., 4,4'-dimethoxytritil; Q is a linker connecting the nitrogen and oxygen, e.g., n-hexyl; R.sub.1 is a nitrogen substituent, e.g., hydrogen; R.sub.2 through R.sub.4 are separately hydrogen or lower alkyl; Y is an atom which is electronegative with respect to carbon, e.g., oxygen or sulfur; X.sub.1 is an atom which is electronegative with respect to carbon, e.g., sulfone; Z is a bond or spacer arm, e.g., ethylsuccinate; and W is a derivatized solid synthesis support capable of linking to Z, e.g., an amino-dirivitized controlled pore glass. The solid support is treated with acid to remove the acid-cleavable hydroxyl protecting group. A protected nucleoside monomer and a weak acid is added forming a linkage, and the unreacted sites on the solid support are capped. An oxidizing reagent is added and the above steps are repeated until polynucleotide chain elongation is complete. The polynucleotide is cleaved from the solid support and then deprotected.
摘要翻译：公开了一种在3'端合成具有氮原子的多核苷酸的方法。方法包括以下步骤。提供具有如下结构的固体支持剂：其中T是酸可裂解的羟基保护基团，例如4,4'-二甲氧基三苯乙酮; Q是连接氮和氧的接头，例如正己基; R1是氮取代基，例如氢; R2至R4分别为氢或低级烷基; Y是相对于碳例如氧或硫具有电负性的原子; X1是相对于碳电负性的原子，例如砜; Z是键或间隔臂，例如乙基琥珀酸盐; 并且W是能够与Z连接的衍生的固体合成载体，例如氨基 - 二硝基化的受控孔玻璃。用酸处理固体支持物以除去酸可裂解的羟基保护基团。加入保护的核苷单体和弱酸形成连接，并将固体载体上的未反应部位加封。加入氧化试剂，并重复上述步骤，直到多核苷酸链延伸完成。多核苷酸从固体支持物上切割，然后去保护。

7. 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。

8. 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。

9. 发明授权

US5258538A 2,3-disubstituted-1,3,2-oxazaphosphacycloalkanes as nucleic acid linking agents 失效
标题翻译： 2,3-二取代-1,3,2-氧杂氮杂环烷烃作为核酸连接剂
公开(公告)号：US5258538A
公开(公告)日：1993-11-02
申请号：US734575
申请日：1991-10-29
申请人： Steven Fung , Sam L. Woo , Lloyd M. Smith
发明人： Steven Fung , Sam L. Woo , Lloyd M. Smith
IPC分类号： A61K47/48 , C07F9/24 , C07F9/6584 , C07H21/00 , C07F9/00
CPC分类号： C07H21/00 , A61K47/48023 , A61K47/48061 , A61K47/48084 , C07F9/2408 , C07F9/65842
摘要： The compounds of the invention include novel linking agents comprising 2-substituted-3-protected-1,3,2-oxazaphosphacycloalkanes and their phosphoramidite precursors. The compounds of the invention further include conjugates of the above linking agents with oligonucleotides and polymer supports. The compounds of the present invention are useful for linking organic moieties, such as fluorescent or chromogenic dyes, to polymer supports and oligonucleotides, particularly single- and double-stranded DNA and RNA fragments.
摘要翻译：本发明的化合物包括包含2-取代-3-保护的1,3,2-氧杂氮杂环烷烃及其亚磷酰胺前体的新型交联剂。本发明的化合物还包括上述连接剂与寡核苷酸和聚合物载体的缀合物。本发明的化合物可用于将有机部分如荧光或显色染料与聚合物载体和寡核苷酸，特别是单链和双链DNA和RNA片段连接。

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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