公开(公告)号：WO1995016910A1

公开(公告)日：1995-06-22

申请号：PCT/US1994012739

申请日：1994-11-04

Applicant: PERKIN-ELMER CORPORATION ,  MADABHUSHI, Ramakrishna, S. ,  MENCHEN, Steven, M. ,  EFCAVITCH, J., William ,  GROSSMAN, Paul, D.

Inventor： PERKIN-ELMER CORPORATION

IPC: G01N27/447

CPC classification number: G01N27/44752 ,  G01N27/44747

Abstract: 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 DEG C to about 50 DEG 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 x 10 to about 1 x 10 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.

Abstract translation: 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）在约6至约6范围内pH范围的水性介质中不存在带电基团 在一个实施方案中，本发明的聚合物选自聚内酰胺类，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。

公开(公告)号：WO1995016911A1

公开(公告)日：1995-06-22

申请号：PCT/US1994013852

申请日：1994-12-06

Applicant: PERKIN-ELMER CORPORATION ,  MADABHUSHI, Ramakrishna, S. ,  MENCHEN, Steven, M. ,  EFCAVITCH, J., William ,  GROSSMAN, Paul, D.

Inventor： PERKIN-ELMER CORPORATION

IPC: G01N27/447

CPC classification number: G01N27/44752 ,  G01N27/44747

Abstract: 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 DEG C to about 50 DEG 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 x 10 to about 1 x 10 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.

Abstract translation: 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）在约6至约6范围内pH范围的水性介质中不存在带电基团 在一个实施方案中，本发明的聚合物选自聚内酰胺类，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。

公开(公告)号：WO1995003066A1

公开(公告)日：1995-02-02

申请号：PCT/US1994008363

申请日：1994-07-25

Applicant: PERKIN-ELMER CORPORATION ,  BOYD, Victoria, L. ,  BOZZINI, Merilisa

Inventor： PERKIN-ELMER CORPORATION

IPC: A61K37/02

CPC classification number: G01N33/6842 ,  C07K1/12 ,  G01N33/6821

Abstract: A method of converting an N-protected amino acid to a corresponding amino acid thiohydantoin is disclosed. The method comprises reacting an N-protected amino acid with a compound represented as follows (OR)(OR')P(=O)X or (OR)(OR')P(=O)-O-P(=O)(OR'')(OR''') wherein R, R', R'', and R''' are each alkyl, aryl or aralkyl groups and X is halogen. The activated N-protected amino acid thus obtained is then reacted with a thiocyanate reagent to generate an N-protected amino acid thiohydantoin.

Abstract translation: 公开了将N-保护的氨基酸转化为相应的氨基酸硫代乙内酰脲的方法。 该方法包括使N-保护的氨基酸与如下所示的化合物（OR）（OR'）P（= O）X或（OR）（OR'）P（= O）-OP（= O） （OR'“）其中R，R'，R”和R“'各自为烷基，芳基或芳烷基，X为卤素。 然后将由此获得的活化的N-保护的氨基酸与硫氰酸酯试剂反应以产生N-保护的氨基酸硫代乙内酰脲。

公开(公告)号：WO1995021378A1

公开(公告)日：1995-08-10

申请号：PCT/US1995001487

申请日：1995-02-06

Applicant: PERKIN-ELMER CORPORATION ,  HING, Paul, A. ,  HOFF, Louis, B. ,  LACHENMEIER, Eric, W. ,  MEAD, Dennis, E. ,  SIMKINS, Barry, A. ,  EFCAVITCH, J., William

Inventor： PERKIN-ELMER CORPORATION

IPC: G01N27/447

CPC classification number: G01N27/44721

Abstract: The present invention relates to a fluorescent polynucleotide analyzer system which utilizes electrophoresis, and in particular, capillary-based electrophoresis. The analyzer system of the present invention is comprised of, in combination, a capillary containing an electrophoretic separation medium, a means for introducing a mixture of fluorescently labeled polynucleotide fragments into the capillary, detection means for detecting the fluorescently labeled polynucleotide fragments, including the simultaneous detection of multiple fluorescent labels, and, means for introducing and removing electrophoretic separation medium from the capillary.

Abstract translation: 本发明涉及利用电泳，特别是毛细管电泳的荧光多核苷酸分析仪系统。 本发明的分析器系统组合包括含有电泳分离介质的毛细管，用于将荧光标记的多核苷酸片段的混合物引入毛细管的装置，用于检测荧光标记的多核苷酸片段的检测装置，包括同时 检测多个荧光标记物，以及用于从毛细管引入和去除电泳分离介质的装置。

公开(公告)号：WO1995020156A1

公开(公告)日：1995-07-27

申请号：PCT/US1995000916

申请日：1995-01-23

Applicant: PERKIN-ELMER CORPORATION ,  WIKTOROWICZ, John

Inventor： PERKIN-ELMER CORPORATION

IPC: G01N27/447

CPC classification number: G01N27/44743 ,  C07K1/26

Abstract: A Capillary Electrophoresis apparatus and method are disclosed which utilize a meltable plug in an end of the capillary tube to selectively pass small ionic contaminants electrophoretically and retain macromolecular analytes against one end of the plug until the plug is melted. When this plug is melted, the analytes and thecontaminants pass through unimpeded. This permits separation of the analytes from the contaminants during electrophoretic separation and enhances instrument resolution.

Abstract translation: 公开了一种毛细管电泳装置和方法，其利用毛细管末端的可熔塞以选择性地通过电泳方式通过小离子污染物，并将大分子分析物保留在塞的一端，直到塞子熔化。 当这种塞子熔化时，分析物和污染物无阻碍地通过。 这样可以在电泳分离期间将分析物与污染物分离，并提高仪器分辨率。

公开(公告)号：WO1995033200A1

公开(公告)日：1995-12-07

申请号：PCT/US1995006465

申请日：1995-05-22

Applicant: PERKIN-ELMER CORPORATION

Inventor： PERKIN-ELMER CORPORATION ,  SCHAWE, Juergen ,  MARGULIES, Marcel

IPC: G01N25/00

CPC classification number: G01K17/00 ,  G01N25/4833 ,  G01N25/4866

Abstract: The invention is directed to a differential analysis method and apparatus (10) wherein a sample (58) and a reference (56) are subjected to an externally applied disturbance, such as a temperature change, in accord with a prescribed function comprising the sum of a linearly changing part and a periodically changing part, and the measured differential signal is processed into real and imaginary components relating, respectively, to the energy storage and energy loss portions of the signal.

Abstract translation: 本发明涉及一种差分分析方法和装置（10），其中样品（58）和参考物（56）根据规定的功能进行外部施加的干扰，例如温度变化，该规定的功能包括 线性变化部分和周期性变化部分，并且所测量的差分信号被分别与信号的能量存储和能量损失部分相关联的实分量和虚分量进行处理。

公开(公告)号：WO1995030139A1

公开(公告)日：1995-11-09

申请号：PCT/US1995004818

申请日：1995-04-19

Applicant: PERKIN-ELMER CORPORATION

Inventor： PERKIN-ELMER CORPORATION ,  WOUDENBERG, Timothy, M. ,  BODNER, Kevin, S. ,  CONNELL, Charles, R. ,  GANZ, Alan, M. ,  MCBRIDE, Lincoln, J. ,  SAVIANO, Paul, G. ,  SHIGEURA, John ,  TRACY, David, H. ,  YOUNG, Eugene, F. ,  LEE, Linda, G.

IPC: G01N21/64

CPC classification number: G01N21/6428 ,  C12Q1/6818 ,  C12Q1/6851 ,  C12Q1/686 ,  G01N21/0303 ,  G01N21/645 ,  G01N2021/6484 ,  C12Q2561/101 ,  C12Q2545/101 ,  C12Q2537/143

Abstract: A system is provided for carrying out real time fluorescence-based measurements of nucleic acid amplification products. In a preferred embodiment of the invention, an excitation beam is focused into a reaction mixture through a surface, the reaction mixture containing: (i) a first fluorescent indicator capable of generating a first fluorescent signal whose intensity is proportional to the amount of an amplification product in the volume of the reaction mixture illuminated by the excitation beam and (ii) a second fluorescent indicator homogeneously distributed throughout the reaction mixture capable of generating a second fluorescent signal proportional to the volume of reaction mixture illuminated by the excitation beam. Preferably, the excitation beam is focused into the reaction mixture by a lens through a portion of a wall of a closed reaction chamber containing the reaction mixture. The same lens is used to collect the first and second fluorescent signals generated by the first and second fluorescent indicators, respectively, in response to the excitation beam. The ratio of the fluorescent intensities of the first and second fluorescent signals provides a stable quantitative indicator of the amount of amplification product synthesized in the course of the amplification reaction.

Abstract translation: 提供了一种用于进行核酸扩增产物的实时基于荧光的测量的系统。 在本发明的优选实施方案中，激发光束通过表面聚焦到反应混合物中，反应混合物包含：（i）第一荧光指示器，其能够产生第一荧光信号，其强度与放大量成比例 由激发束照射的反应混合物的体积的产物和（ii）均匀分布在整个反应混合物中的第二荧光指示剂，其能够产生与由激发束照射的反应混合物的体积成比例的第二荧光信号。 优选地，激发光束通过透镜通过包含反应混合物的封闭反应室的壁的一部分聚焦到反应混合物中。 相同的透镜被用于响应于激发光束分别收集由第一和第二荧光指示器产生的第一和第二荧光信号。 第一和第二荧光信号的荧光强度比提供了在扩增反应过程中合成的扩增产物的量的稳定的定量指标。

公开(公告)号：WO1995014921A1

公开(公告)日：1995-06-01

申请号：PCT/US1994011889

申请日：1994-10-18

Applicant: PERKIN-ELMER CORPORATION ,  JOHNSON, Ben, F. ,  MENCHEN, Steven, M. ,  BLOCH, Will

Inventor： PERKIN-ELMER CORPORATION

IPC: G01N27/447

CPC classification number: G01N27/44743 ,  C12N15/101 ,  C12Q1/6869 ,  G01N27/44747

Abstract: A capillary electrophoresis system and method of electrokinetically loading a capillary electrophoresis sample into a separation medium in a capillary tube in which an entangled polymer matrix is formed having the sample embedded therein. The matrix has a mesh size effective to retard movement of macromolecules such as DNA sequencing templates through the matrix when an electric field is applied across the matrix. The entangled polymer matrix is formed by a linear polymer having a molecular weight of at least 20K Daltons.

Abstract translation: 毛细管电泳系统和将毛细管电泳样品电动加载到毛细管中的分离介质中的毛细管电泳系统，其中形成缠绕聚合物基质的样品嵌入其中。 当电场施加在基质上时，该基质具有有效的阻止大分子如DNA测序模板通过基质的移动的筛孔。 缠结聚合物基质由分子量至少为20K道尔顿的线性聚合物形成。

公开(公告)号：WO1996015270A1

公开(公告)日：1996-05-23

申请号：PCT/US1995014882

申请日：1995-11-15

Applicant: PERKIN-ELMER CORPORATION

Inventor： PERKIN-ELMER CORPORATION ,  LIVAK, Kenneth, J. ,  FLOOD, Susan, J., A. ,  MARAMARO, Jeffrey ,  MULLAH, Khairuzzaman, Bashar

IPC: C12Q01/68

CPC classification number: C12Q1/6818 ,  C12Q1/6834 ,  C12Q1/6837 ,  C12Q2565/519 ,  C12Q2563/107 ,  C12Q2561/101 ,  C12Q2525/185 ,  C12Q2525/301 ,  C12Q2565/1015 ,  C12Q2565/107

Abstract: An oligonucleotide probe is provided which includes a fluorescent reporter molecule and a quencher molecule capable of quenching the fluorescence of the reporter molecule. The oligonucleotide probe is constructed such that the probe exists in at least one single-stranded conformation when unhybridized where the quencher molecule is near enough to the reporter molecule to quench the fluorescence of the reporter molecule. The oligonucleotide probe also exists in at least one conformation when hybridized to a target polynucloetide where the quencher molecule is not positioned close enough to the reporter molecule to quench the fluorescence of the reporter molecule. By adopting these hybridized and unhybridized conformations, the reporter molecule and quencher molecule on the probe exhibit different fluorescence signal intensities when the probe is hybridized and unhybridized. As a result, it is possible to determine whether the probe is hybridized or unhybridized based on a change in the fluorescence intensity of the reporter molecule, the quencher molecule, or a combination thereof. In addition, because the probe can be designed such that the quencher molecule quenches the reporter molecule when the probe is not hybridized, the probe can be designed such that the reporter molecule exhibits limited fluorescence until the probe is either hybridized or digested.

Abstract translation: 提供了一种寡核苷酸探针，其包括荧光报道分子和能够猝灭报道分子荧光的猝灭剂分子。 构建寡核苷酸探针，使得探针在未杂交的情况下以至少一个单链构象存在，其中猝灭剂分子足够接近报道分子以淬灭报道分子的荧光。 当寡聚核苷酸探针与目标多烯苏胺杂交时，寡核苷酸探针也存在至少一个构象，其中猝灭剂分子不能靠近报告分子，以猝灭报道分子的荧光。 通过采用这些杂交和非杂交构象，当探针杂交和未杂交时，探针上的报道分子和猝灭剂分子表现出不同的荧光信号强度。 结果，可以基于报道分子，猝灭剂分子或其组合的荧光强度的变化来确定探针是杂交还是未杂交。 此外，因为探针可以被设计成使得当探针不杂交时，猝灭剂分子淬灭报告分子，所以探针可被设计成使得报告分子在荧光探针杂交或消化之前显示有限的荧光。

公开(公告)号：WO1996006190A3

公开(公告)日：1996-02-29

申请号：PCT/US1995010603

申请日：1995-08-17

Applicant: PERKIN-ELMER CORPORATION

Inventor： PERKIN-ELMER CORPORATION ,  EGGERDING, Faye

IPC: C12Q01/68