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31. 发明授权

US5795729A Reductive, energy-transfer fluorogenic probes 失效
标题翻译：还原性，能量转移荧光探针
公开(公告)号：US5795729A
公开(公告)日：1998-08-18
申请号：US597018
申请日：1996-02-05
申请人： Linda G. Lee
发明人： Linda G. Lee
IPC分类号： G01N33/542 , C12Q1/37 , C12Q1/00 , C12Q1/26 , G01N33/53
CPC分类号： G01N33/542 , Y10S435/968
摘要： An energy transfer fluorescent probe for detecting a reagent is provided which includes a fluorescent reporter molecule and a quencher molecule positioned on the probe relative to the reporter molecule such that the quencher molecule quenches the fluorescence of the reporter molecule when in a first state, the quencher molecule being converted by the reagent to a second state which has a reduced ability to quench the reporter molecule. Examples of conversions of the quencher molecule from a first state to a second state include reductions, oxidations, hydrolyses, phosphate cleavages, and the conversion of amides to amines. In one embodiment, the quencher molecule is a substrate for an enzyme which converts the quencher from a first state to a second state. For example, the enzyme may be an reductase, an oxidase, hydrolytic, a peptidase or a phosphorylase. The probe is used to fluorescently detect a reagent in a sample. According to the method, the probe is contacted with the sample containing the reagent wherein the quencher molecule is converted by the reagent from a first state which is able to quench the fluorescence of the reporter molecule to a second state which has a reduced ability to quench the reporter molecule. The reagent is detected by monitoring the fluorescence intensity of the reporter molecule as the quencher molecule is converted from the first state to the second state by the reagent.
摘要翻译：提供了一种用于检测试剂的能量转移荧光探针，其包含相对于报道分子位于探针上的荧光报道分子和猝灭剂分子，使得猝灭剂分子在处于第一状态时淬灭报道分子的荧光，淬灭剂分子由试剂转化成具有降低报告分子淬灭能力的第二状态。猝灭剂分子从第一种状态转化为第二种状态的实例包括还原，氧化，水解，磷酸盐裂解和酰胺转化为胺。在一个实施方案中，猝灭剂分子是用于将猝灭剂从第一状态转化为第二状态的酶的底物。例如，酶可以是还原酶，氧化酶，水解酶，肽酶或磷酸化酶。探针用于荧光检测样品中的试剂。根据该方法，将探针与含有试剂的样品接触，其中猝灭剂分子由试剂转化为能够将报道分子的荧光猝灭的第一状态到具有降低的淬灭能力的第二状态记者分子。当猝灭剂分子由试剂从第一状态转变为第二状态时，通过监测报道分子的荧光强度来检测试剂。

32. 发明申请

US20120172583A1 Oligonucleotides and Analogs Labeled with Energy Transfer Dyes 审中-公开
标题翻译：标记能量转移染料的寡核苷酸和类似物
公开(公告)号：US20120172583A1
公开(公告)日：2012-07-05
申请号：US12882034
申请日：2010-09-14
申请人： Linda G. Lee , Sandra L. Spurgeon , Barnett B. Rosenblum
发明人： Linda G. Lee , Sandra L. Spurgeon , Barnett B. Rosenblum
IPC分类号： C07H21/00 , C07D311/90 , C07D311/88 , C07D407/12 , C07D491/22 , C07D491/16
CPC分类号： C07H21/04 , C07D311/90
摘要： Novel linkers for linking a donor dye to an acceptor dye in an energy transfer fluorescent dye are provided. These linkers facilitate the efficient transfer of energy between a donor and acceptor dye in an energy transfer dye. One of these linkers for linking a donor dye to an acceptor dye in an energy transfer fluorescent dye has the general structure R21Z1C(O)R22R28 where R21 is a C1-5 alkyl attached to the donor dye, C(O) is a carbonyl group, Z1 is either NH, sulfur or oxygen, R22 is a substituent which includes an alkene, diene, alkyne, a five and six membered ring having at least one unsaturated bond or a fused ring structure which is attached to the carbonyl carbon, and R28 includes a functional group which attaches the linker to the acceptor dye.
摘要翻译：提供了用于将供体染料与能量转移荧光染料中的受体染料连接的新型接头。这些接头促进能量转移染料中供体和受体染料之间的能量的有效转移。用于将供体染料与能量转移荧光染料中的受体染料连接的这些接头之一具有通式结构R21Z1C（O）R22R28，其中R21是与供体染料连接的C 1-5烷基，C（O）是羰基 Z1是NH，硫或氧，R22是包含烯烃，二烯烃，炔烃，具有至少一个不饱和键的五元和六元环或连接到羰基碳上的稠环结构的取代基，R28 包括将接头连接到受体染料的官能团。

33. 发明授权

US08211673B2 Composition and method for sequencing nucleic acid 有权
标题翻译：核酸测序的组成和方法
公开(公告)号：US08211673B2
公开(公告)日：2012-07-03
申请号：US12365140
申请日：2009-02-03
申请人： Linda G. Lee
发明人： Linda G. Lee
IPC分类号： C12P19/34 , C12Q1/68
CPC分类号： C12Q1/6806 , C12Q1/6848 , C12Q1/6853 , C12Q2521/325 , C12Q2525/125 , C12Q2535/101
摘要： A composition for sequencing DNA is provided and comprises a nuclease and a nuclease-resistant sequencing primer. A method of preparing DNA for sequencing and a method of sequencing DNA are also provided. The method of sequencing DNA can comprise contacting amplification reaction products with the composition under conditions in which excess amplification primer is degraded by the nuclease and the nuclease-resistant sequencing primer is essentially non-degraded.
摘要翻译：提供了用于测序DNA的组合物，并且包含核酸酶和核酸酶抗性测序引物。还提供了制备用于测序的DNA的方法和测序DNA的方法。测序DNA的方法可以包括在扩增引物被核酸酶降解的条件下将扩增反应产物与组合物接触，并且核酸酶抗性测序引物基本上是未降解的。

34. 发明授权

US07955864B2 Device and method for making discrete volumes of a first fluid in contact with a second fluid, which are immiscible with each other 有权
标题翻译：使与第二流体接触的第一流体的离散体积彼此不混溶的装置和方法
公开(公告)号：US07955864B2
公开(公告)日：2011-06-07
申请号：US11508044
申请日：2006-08-22
申请人： David M. Cox , Willy Wiyatno , Mark F. Oldham , James C. Nurse , Douglas P. Greiner , Sam L. Woo , Richard T. Reel , Dennis A. Lehto , Linda G. Lee , Janusz B. Wojtowicz
发明人： David M. Cox , Willy Wiyatno , Mark F. Oldham , James C. Nurse , Douglas P. Greiner , Sam L. Woo , Richard T. Reel , Dennis A. Lehto , Linda G. Lee , Janusz B. Wojtowicz
IPC分类号： G01N1/00
CPC分类号： C12Q1/6869 , B01J2219/00353 , B01J2219/0036 , B01J2219/00364 , B01J2219/00608 , B01J2219/0061 , B01J2219/00612 , B01J2219/00619 , B01J2219/00626 , B01J2219/00637 , B01J2219/00653 , B01J2219/00657 , B01J2219/00659 , B01L3/0293 , B01L3/502715 , B01L3/502784 , B01L7/52 , B01L2200/0673 , B01L2200/10 , B01L2300/0864 , B01L2300/0867 , B01L2400/0421 , B01L2400/0487 , C12Q1/6806 , C12Q1/6874 , F15C5/00 , F16K99/0001 , F16K99/0011 , F16K99/0013 , F16K2099/0084 , G01N1/14 , G01N27/44743 , G01N27/44769 , G01N35/08 , Y10T137/0318 , Y10T137/4259 , Y10T137/85978 , Y10T137/85986 , Y10T137/86863 , Y10T436/2575 , C12Q2535/101
摘要： Various embodiments described in the application relate to an apparatus, system, and method for generating, within a conduit, discrete volumes of one or more fluids that are immiscible with a second fluid. The discrete volumes can be used for biochemical or molecular biology procedures involving small volumes, for example, microliter-sized volumes, nanoliter-sized volumes, or smaller. The system can comprise an apparatus comprising at least one conduit operatively connected to one or more pumps for providing discrete volumes separated from one another by a fluid that is immiscible with the fluid(s) of the discrete volumes, for example, aqueous immiscible-fluid-discrete volumes separated by an oil.
摘要翻译：在本申请中描述的各种实施例涉及用于在管道内产生与第二流体不混溶的一种或多种流体的离散体积的装置，系统和方法。离散体积可用于涉及小体积的生物化学或分子生物学过程，例如微升体积，纳升级体积或更小体积。该系统可以包括一种装置，该装置包括至少一个导管，其操作地连接到一个或多个泵，用于提供通过与离散体积的流体不混溶的流体彼此分离的离散体积，例如水不混溶流体 - 油分离体积。

35. 发明授权

US07595162B2 Methods of labeling polynucleotides with energy transfer dyes 失效
标题翻译：用能量转移染料标记多核苷酸的方法
公开(公告)号：US07595162B2
公开(公告)日：2009-09-29
申请号：US11618667
申请日：2006-12-29
申请人： Linda G. Lee , Sandra L. Spurgeon , Barnett Rosenblum
发明人： Linda G. Lee , Sandra L. Spurgeon , Barnett Rosenblum
IPC分类号： C12P19/34 , C07H21/00 , C07H21/02
CPC分类号： C07H21/04 , C07D311/82 , C07D311/90 , C07D405/12 , C07D405/14 , C07H21/00 , C09B11/04 , C09B11/24 , C09B23/083 , C09B69/00 , C12Q1/6818 , C12Q1/6869 , C12Q2563/107 , C12Q2535/101 , C12Q2565/1015
摘要： Novel linkers for linking a donor dye to an acceptor dye in an energy transfer fluorescent dye are provided. These linkers faciliate the efficient transfer of energy between a donor and acceptor dye in an energy transfer dye. One of these linkers for linking a donor dye to an acceptor dye in an energy transfer fluorescent dye has the general structure R21Z1C(O)R22R28 where R21 is a C1-5 alkyl attached to the donor dye, C(O) is a carbonyl group, Z1 is either NH, sulfur or oxygen, R22 is a substituent which includes an alkene, diene, alkyne, a five and six membered ring having at least one unsaturated bond or a fused ring structure which is attached to the carbonyl carbon, and R28 includes a functional group which attaches the linker to the acceptor dye.
摘要翻译：提供了用于将供体染料与能量转移荧光染料中的受体染料连接的新型接头。这些连接器有助于在能量转移染料中的供体和受体染料之间的能量的有效转移。用于将供体染料与能量转移荧光染料中的受体染料连接的这些接头之一具有通式结构R21Z1C（O）R22R28，其中R21是与供体染料连接的C 1-5烷基，C（O）是羰基 Z1是NH，硫或氧，R22是包含烯烃，二烯烃，炔烃，具有至少一个不饱和键的五元和六元环或连接到羰基碳上的稠环结构的取代基，R28 包括将接头连接到受体染料的官能团。

36. 发明申请

US20090031506A1 UV Excitable Fluorescent Energy Transfer Dyes 有权
标题翻译：紫外线可激发荧光能量转移染料
公开(公告)号：US20090031506A1
公开(公告)日：2009-02-05
申请号：US12126813
申请日：2008-05-23
申请人： Linda G. Lee
发明人： Linda G. Lee
IPC分类号： C09K11/06 , C09B57/02
CPC分类号： C12Q1/6869 , C07H19/10 , C07H19/20 , C07H21/00 , C07H21/04 , C09B11/24 , C12Q1/6818 , C12Q2563/107
摘要： Novel energy transfer dyes which can be used with shorter wavelength light sources are provided. These dyes include a donor dye with an absorption maxima at a wavelength between about 250 to 450 nm and an acceptor dye which is capable of absorbing energy emitted from the donor dye. One of the energy transfer dyes has a donor dye which is a member of a class of dyes having a coumarin or pyrene ring structure and an acceptor dye which is capable of absorbing energy emitted from the donor dye, wherein the donor dye has an absorption maxima between about 250 and 450 nm and the acceptor dye has an emission maxima at a wavelength greater than about 500 nm.
摘要翻译：提供了可用于较短波长光源的新型能量转移染料。这些染料包括在约250至450nm之间的波长处具有最大吸收的供体染料和能够吸收从供体染料发射的能量的受体染料。能量转移染料之一具有供体染料，其是具有香豆素或芘环结构的一类染料的成员和能够吸收从供体染料发射的能量的受体染料，其中供体染料具有吸收最大值在约250和450nm之间，受体染料具有波长大于约500nm的发射最大值。

37. 发明申请

US20080268509A1 METHODS OF LABELING POLYNUCLEOTIDES WITH ENERGY TRANSFER DYES 失效
标题翻译：用能量转移标记标记多核苷酸的方法
公开(公告)号：US20080268509A1
公开(公告)日：2008-10-30
申请号：US11618667
申请日：2006-12-29
申请人： Linda G. Lee , Sandra L. Spurgeon , Barnett Rosenblum
发明人： Linda G. Lee , Sandra L. Spurgeon , Barnett Rosenblum
IPC分类号： C12P19/34
CPC分类号： C07H21/04 , C07D311/82 , C07D311/90 , C07D405/12 , C07D405/14 , C07H21/00 , C09B11/04 , C09B11/24 , C09B23/083 , C09B69/00 , C12Q1/6818 , C12Q1/6869 , C12Q2563/107 , C12Q2535/101 , C12Q2565/1015
摘要： Novel linkers for linking a donor dye to an acceptor dye in an energy transfer fluorescent dye are provided. These linkers faciliate the efficient transfer of energy between a donor and acceptor dye in an energy transfer dye. One of these linkers for linking a donor dye to an acceptor dye in an energy transfer fluorescent dye has the general structure R21Z1C(O)R22R28 where R21 is a C1-5 alkyl attached to the donor dye, C(O) is a carbonyl group, Z1 is either NH, sulfur or oxygen, R22 is a substituent which includes an alkene, diene, alkyne, a five and six membered ring having at least one unsaturated bond or a fused ring structure which is attached to the carbonyl carbon, and R28 includes a functional group which attaches the linker to the acceptor dye.
摘要翻译：提供了用于将供体染料与能量转移荧光染料中的受体染料连接的新型接头。这些连接器有助于在能量转移染料中的供体和受体染料之间的能量的有效转移。用于将供体染料与能量转移荧光染料中的受体染料连接的这些接头之一具有通式结构R 21 R 1 C（O）R 22 其中R 21是与供体染料连接的C 1-5烷基，C（O）是羰基（C 1 -C 4）烷基，基团Z 1是NH，硫或氧，R 22是包括烯烃，二烯烃，炔烃，具有至少一个碳原子的五元和六元环的取代基不饱和键或与羰基碳连接的稠合环结构，R 28包括将接头连接于受体染料的官能团。

38. 发明授权

US06649769B2 Atropisomers of asymmetric xanthene fluorescent dyes and methods of DNA sequencing and fragment analysis 失效
标题翻译：不对称呫吨荧光染料的倾向异构体和DNA测序和片段分析方法
公开(公告)号：US06649769B2
公开(公告)日：2003-11-18
申请号：US10227058
申请日：2002-08-21
申请人： Linda G. Lee , Meng C. Taing , Barnett B. Rosenblum
发明人： Linda G. Lee , Meng C. Taing , Barnett B. Rosenblum
IPC分类号： C07D40304
CPC分类号： C07H19/10 , C07H19/20 , C07H21/00 , C07H21/04 , C09B11/08 , C09B11/24 , G01N33/533
摘要： Atropisomeric energy-transfer dye compounds are disclosed. A variety of molecular biology applications utilize atropisomeric xanthene fluorescent dyes as labels for substrates such as nucleotides, nucleosides, polynucleotides, polypeptides and carbohydrates. Methods include DNA sequencing, DNA fragment analysis, PCR, SNP analysis, oligonucleotide ligation, amplification, minisequencing, and primer extension.
摘要翻译：公开了异转移能量转移染料化合物。各种分子生物学应用利用无异构呫吨荧光染料作为底物的标记，例如核苷酸，核苷，多核苷酸，多肽和碳水化合物。方法包括DNA测序，DNA片段分析，PCR，SNP分析，寡核苷酸连接，扩增，minisequencing和引物延伸。

39. 发明授权

US06316610B1 Labelled oligonucleotides synthesized on solid-supports 有权
公开(公告)号：US06316610B1
公开(公告)日：2001-11-13
申请号：US09813378
申请日：2001-03-20
申请人： Linda G. Lee , Khairuzzaman B. Mullah , Barnett B. Rosenblum
发明人： Linda G. Lee , Khairuzzaman B. Mullah , Barnett B. Rosenblum
IPC分类号： C12Q168
摘要： Methods and compositions to label oligonucleotides and analogs directly on a solid-support having the structure where S is a solid-support, A is a cleavable linker, X is a moiety with three or more attachment sites, L is a label, Y is a nucleophile, i.e. O, NH, NR or S, and P1 is an acid cleavable protecting group are provided. The labelled solid-support is reacted in a cyclical fashion to synthesize a labelled oligonucleotide on a solid-support in the 5′ to 3′ direction, having the structure: Labelled oligonucleotides are also synthesized by reacting: (i) a label reagent bearing functionality consisting of carboxylic acid, sulfonic acid, phosphonic acid, or phosphoric acid, (ii) an oligonucleotide on solid support with nucleophilic functionality, and (iii) a coupling reagent, whereby an ester, amide, thioester, sulfonamide, sulfonate, phosphonate, phosphoramidate, phosphorothioate, or phosphate bond is formed. The labelling reaction may be conducted at label sites including the 5′ terminus, the 3′ terminus, a nucleobase, an internucleotide linkage, a sugar, amino, sulfide, hydroxyl, and carboxyl.

40. 发明授权

US6080868A Nitro-substituted non-fluorescent asymmetric cyanine dye compounds 失效
标题翻译：硝基取代的非荧光不对称花青染料化合物
公开(公告)号：US6080868A
公开(公告)日：2000-06-27
申请号：US12525
申请日：1998-01-23
申请人： Linda G. Lee , Ronald J. Graham , Khairuzzaman B. Mullah , Francis T. Haxo
发明人： Linda G. Lee , Ronald J. Graham , Khairuzzaman B. Mullah , Francis T. Haxo
IPC分类号： G01N33/58 , C07D413/06 , C07D417/06 , C07D421/06 , C07D513/04 , C07H19/04 , C07H21/00 , C09B23/00 , C09B23/02 , C12N15/09 , C12Q1/68 , G01N21/78 , C07D293/00
CPC分类号： C07H21/00 , C09B23/02
摘要： The invention provides an asymmetric cyanine dye compound having the structure ##STR1## including substituted forms thereof, wherein, at least one of R.sub.1 and R.sub.2 is linking group, X is O, S, or Se, and n ranges from 0 to 2. The invention further provides reporter-quencher dye pairs comprising the asymmetric cyanine dyes, dye-labeled polynucleotides incorporating the asymmetric cyanine dyes, and hybridization detection methods utilizing the dye-labeled polynucleotides.
摘要翻译：本发明提供具有包括其取代形式的结构的不对称花青染料化合物，其中R 1和R 2中的至少一个是连接基团，X是O，S或Se，n在0至2的范围内。本发明进一步提供包含不对称花青染料，掺入不对称花青染料的染料标记的多核苷酸和利用染料标记的多核苷酸的杂交检测方法的报道猝灭剂染料对。

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