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1. 发明公开

EP0884394A3 Apparatus for assembly of an in situ PCR sample and reagent fluid containment system on a glass slide 失效
标题翻译：用于在载玻片上组装原位PCR样品和试剂流体容纳系统的装置
公开(公告)号：EP0884394A3
公开(公告)日：1998-12-23
申请号：EP98115093.1
申请日：1993-09-30
申请人： The Perkin Elmer Corporation
发明人： Atwood, John G. , Haff, Larry
IPC分类号： C12Q1/68 , G02B21/34 , B01L7/00
CPC分类号： G01N1/312 , B01J19/0046 , B01J2219/00722 , B01L3/508 , B01L7/52 , B01L9/52 , B01L2200/0684 , B01L2200/0689 , B01L2300/041 , B01L2300/0822 , B01L2300/1827 , B01L2300/185 , C12Q1/6841 , C12Q1/686 , C40B40/06 , Y10S435/809 , C12Q2543/101
摘要： An apparatus (80) for assembly of an in situ PCR sample and reagent fluid containment system on a glass slide (14), wherein said containment system comprises a cover member (16) and a retaining assembly (20,22, 28), said apparatus comprising: a base (45); a support platform (42) mounted on said base, said support platform having a generally horizontal top surface for receiving and supporting components of said retaining assembly and said slide in a predetermined spaced relation; an elongate compression arm (84) movably supported from said base at a position spaced from said platform, said arm being operative to compress said glass slide and said retaining assembly together in close contact; and a retaining assembly installation mechanism (86) positioned so as to install said retaining assembly onto said slide (14) while said compression arm (84) is compressing said glass slide (14) and said retaining assembly together.
摘要翻译：一种用于在玻璃载玻片（14）上组装原位PCR样品和试剂流体容纳系统的设备（80），其中所述容纳系统包括盖构件（16）和保持组件（20,22,28），所述设备，包括：基座（45）; 安装在所述基座上的支撑平台（42），所述支撑平台具有大致水平的顶表面，用于以预定的间隔关系接收和支撑所述保持组件和所述滑块的部件; 在与所述平台间隔开的位置处从所述基部可移动地支撑的细长压缩臂（84），所述臂可操作以将所述载玻片和所述保持组件紧密接触地压缩在一起; 和保持组件安装机构（86），当所述压缩臂（84）将所述玻璃滑块（14）和所述保持组件压缩在一起时，所述保持组件安装机构（86）定位成将所述保持组件安装到所述滑块（14）上。

2. 发明公开

EP0884394A2 Apparatus for assembly of an in situ PCR sample and reagent fluid containment system on a glass slide 失效
标题翻译：装置用于在薄片原位PCR构建一个样品和试剂接收系统，用于
公开(公告)号：EP0884394A2
公开(公告)日：1998-12-16
申请号：EP98115093.1
申请日：1993-09-30
申请人： The Perkin Elmer Corporation
发明人： Atwood, John G. , Haff, Larry
IPC分类号： C12Q1/68 , G02B21/34 , B01L7/00
CPC分类号： G01N1/312 , B01J19/0046 , B01J2219/00722 , B01L3/508 , B01L7/52 , B01L9/52 , B01L2200/0684 , B01L2200/0689 , B01L2300/041 , B01L2300/0822 , B01L2300/1827 , B01L2300/185 , C12Q1/6841 , C12Q1/686 , C40B40/06 , Y10S435/809 , C12Q2543/101
摘要： An apparatus (80) for assembly of an in situ PCR sample and reagent fluid containment system on a glass slide (14), wherein said containment system comprises a cover member (16) and a retaining assembly (20,22, 28), said apparatus comprising: a base (45); a support platform (42) mounted on said base, said support platform having a generally horizontal top surface for receiving and supporting components of said retaining assembly and said slide in a predetermined spaced relation; an elongate compression arm (84) movably supported from said base at a position spaced from said platform, said arm being operative to compress said glass slide and said retaining assembly together in close contact; and a retaining assembly installation mechanism (86) positioned so as to install said retaining assembly onto said slide (14) while said compression arm (84) is compressing said glass slide (14) and said retaining assembly together.
摘要翻译：在载玻片上的装置（80），用于的组件原位PCR的样品和试剂的流体容纳系统（14）worin所述容纳系统包括盖构件（16）和一个保持组件（20,22，28），所述装置，包括：一个基座（45）; 支撑平台（42）安装在基座说，所说的基因的反弹水平顶面支撑平台，用于接收和在预定的间隔关系支撑所述保持组件和所述滑动件的部件; 伸长在从所述平台隔开的位置从所述基站可移动地支撑压缩臂（84），所述臂被操作以压缩所述载玻片和所述紧密接触在一起保持组件; 和一个保持组件安装机构（86），以便安装定位在所述保持组件到所述滑动件（14）而压缩所述臂（84）压缩所述载玻片（14）和所述保持组件在一起。

3. 发明公开

EP0686699A3 Apparatus and method for determining the concentration of the target nucleic acid in PCR 失效
标题翻译：装置和方法用于确定在PCR的靶核酸的浓度
公开(公告)号：EP0686699A3
公开(公告)日：1996-04-17
申请号：EP95108649.5
申请日：1995-06-06
申请人： THE PERKIN-ELMER CORPORATION
发明人： Atwood, John G.
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6816 , C12Q1/6851
摘要： Apparatus and method for determining an unknown starting molar concentration of target nucleic acid molecules at the beginning of a polymerase chain reaction in a sample reaction mixture containing suitable buffers, two complementary kinds of oligonucleotide primers, a molar excess of four kinds of nucleoside triphosphates, a DNA polymerase, and the unknown starting molar concentration of target nucleic acid molecules, wherein the two kinds of primers are provided in a known concentration (Cp o ), the method including the steps of adding a reporter molecule which does not significantly interfere with the reaction and does not emit a strong signal in the absence of double-stranded DNA, but makes the dsDNA created by the reaction detectable and measurable at least once each cycle, thermal cycling one or more standard reaction mixtures which are substantially the same in every respect except they have known starting molar concentration of target nucleic acid molecules to obtain one or more growth curves with known starting molar concentrations of target nucleic acid molecules, exciting the reaction mixture during at least the extension portion of each cycle, detecting and measuring the intensity of the signal during at least the extension portion of each of the cycles, converting said intensity to molar concentration values of dsDNA and storing the molar concentration values for each of the extension portions of each of the cycles, generating a measured curve of molar concentration of dsDNA versus cycle number from the stored concentration values, using successive approximations, determining values of e v , e s , a, C z (n) and A z (n) which provide a best fit of the measured curve to the one or more known growth curves according to the following relation: where: C n+1 and C n are the DNA template molar concentrations at the end of the n and n + 1 cycle extension periods;
e v is in theory the template's probability of survival to the next cycle; e s is in theory the template's probability of being synthesized; C o is the molar concentration of starting target nucleic acid molecules; Cp o is the starting primer molar concentration; a is in theory the primer/complementary strand competition factor; C z is molar concentration of the polymerase enzyme; A z is specific activity of the polymerase; t z is extension time in seconds; L t is length of template in bases; Lp is length of primers in bases; and
calculating the molar concentration of starting nucleic acid of the unknown samples by fixing e v , e s , a, C z (n), and A z (n), and adjusting the starting concentration C o to get a best fit to the measured growth curve of each unknown sample.

4. 发明授权

EP0611598B1 In situ PCR amplification system 失效
标题翻译：在原位PCR扩增程序系统
公开(公告)号：EP0611598B1
公开(公告)日：1999-03-03
申请号：EP93115813.3
申请日：1993-09-30
申请人： THE PERKIN-ELMER CORPORATION
发明人： Atwood, John G. , Haff, Larry
IPC分类号： B01L7/00 , B01L3/00 , B01J15/00
CPC分类号： G01N1/312 , B01J19/0046 , B01J2219/00722 , B01L3/508 , B01L7/52 , B01L9/52 , B01L2200/0684 , B01L2200/0689 , B01L2300/041 , B01L2300/0822 , B01L2300/1827 , B01L2300/185 , C12Q1/6841 , C12Q1/686 , C40B40/06 , Y10S435/809 , C12Q2543/101

5. 发明公开

EP0611598A3 In situ PCR amplification system 失效
标题翻译：原位PCR扩增过程系统。
公开(公告)号：EP0611598A3
公开(公告)日：1995-02-01
申请号：EP93115813.3
申请日：1993-09-30
申请人： THE PERKIN-ELMER CORPORATION
发明人： Atwood, John G. , Haff, Larry
IPC分类号： B01L7/00 , B01L3/00 , B01J15/00
CPC分类号： G01N1/312 , B01J19/0046 , B01J2219/00722 , B01L3/508 , B01L7/52 , B01L9/52 , B01L2200/0684 , B01L2200/0689 , B01L2300/041 , B01L2300/0822 , B01L2300/1827 , B01L2300/185 , C12Q1/6841 , C12Q1/686 , C40B40/06 , Y10S435/809 , C12Q2543/101
摘要： A complete in situ PCR system for amplification of nucleic acids contained in a prepared cell or tissue sample. The containment system for the PCR sample comprises a glass microscope slide (14), a specimen sample (12) containing the target nucleic acid sequence mounted on the slide, a flexible plastic cover over the sample (16), and a retaining assembly (18) fastened to the slide and to the cover to retain and seal a reaction mixture against the sample during thermal cycling. The retaining assembly includes a rigid ring (20) on a rim portion of the cover, a cross beam (22) having spaced parallel rails (24) joined by opposite flat ends, and a pair of clips (28) which are pressed over the ends and opposite sides of the slide to fasten the cross beam and cover to the slide.

6. 发明公开

EP0611598A2 In situ PCR amplification system 失效
标题翻译：原位PCR Vermehrungsverfahren系统。
公开(公告)号：EP0611598A2
公开(公告)日：1994-08-24
申请号：EP93115813.3
申请日：1993-09-30
申请人： THE PERKIN-ELMER CORPORATION
发明人： Atwood, John G. , Haff, Larry
IPC分类号： B01L7/00 , B01L3/00 , B01J15/00
CPC分类号： G01N1/312 , B01J19/0046 , B01J2219/00722 , B01L3/508 , B01L7/52 , B01L9/52 , B01L2200/0684 , B01L2200/0689 , B01L2300/041 , B01L2300/0822 , B01L2300/1827 , B01L2300/185 , C12Q1/6841 , C12Q1/686 , C40B40/06 , Y10S435/809 , C12Q2543/101
摘要： A complete in situ PCR system for amplification of nucleic acids contained in a prepared cell or tissue sample. The containment system for the PCR sample comprises a glass microscope slide (14), a specimen sample (12) containing the target nucleic acid sequence mounted on the slide, a flexible plastic cover over the sample (16), and a retaining assembly (18) fastened to the slide and to the cover to retain and seal a reaction mixture against the sample during thermal cycling.
The retaining assembly includes a rigid ring (20) on a rim portion of the cover, a cross beam (22) having spaced parallel rails (24) joined by opposite flat ends, and a pair of clips (28) which are pressed over the ends and opposite sides of the slide to fasten the cross beam and cover to the slide.
摘要翻译：用于扩增制备的细胞或组织样品中所含核酸的完整原位PCR系统。用于PCR样品的容纳系统包括玻璃显微镜载玻片，包含安装在载玻片上的靶核酸序列的样本样品，样品上的柔性塑料盖，以及固定到载玻片和盖以保持和在热循环期间将反应混合物密封在样品上。保持组件包括在盖的边缘部分上的刚性环，具有由相对的平坦端连接的间隔开的平行轨道的横梁以及压在滑块的端部和相对侧上的一对夹子，以将横梁并覆盖到幻灯片。

7. 发明授权

EP0059836B1 Optical beam splitter 失效
标题翻译：光束分离器
公开(公告)号：EP0059836B1
公开(公告)日：1985-08-21
申请号：EP82100648.3
申请日：1982-01-29
申请人： THE PERKIN-ELMER CORPORATION
发明人： Helms, Charles C. , Atwood, John G.
IPC分类号： G02B27/14 , G01J3/42
CPC分类号： G02B27/143 , G01J1/36

8. 发明公开

EP0686699A2 Apparatus and method for determining the concentration of the target nucleic acid in PCR 失效
标题翻译： PCR和PCR方法的最佳方法
公开(公告)号：EP0686699A2
公开(公告)日：1995-12-13
申请号：EP95108649.5
申请日：1995-06-06
申请人： THE PERKIN-ELMER CORPORATION
发明人： Atwood, John G.
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6816 , C12Q1/6851
摘要： Apparatus and method for determining an unknown starting molar concentration of target nucleic acid molecules at the beginning of a polymerase chain reaction in a sample reaction mixture containing suitable buffers, two complementary kinds of oligonucleotide primers, a molar excess of four kinds of nucleoside triphosphates, a DNA polymerase, and the unknown starting molar concentration of target nucleic acid molecules, wherein the two kinds of primers are provided in a known concentration (Cp o ), the method including the steps of adding a reporter molecule which does not significantly interfere with the reaction and does not emit a strong signal in the absence of double-stranded DNA, but makes the dsDNA created by the reaction detectable and measurable at least once each cycle, thermal cycling one or more standard reaction mixtures which are substantially the same in every respect except they have known starting molar concentration of target nucleic acid molecules to obtain one or more growth curves with known starting molar concentrations of target nucleic acid molecules, exciting the reaction mixture during at least the extension portion of each cycle, detecting and measuring the intensity of the signal during at least the extension portion of each of the cycles, converting said intensity to molar concentration values of dsDNA and storing the molar concentration values for each of the extension portions of each of the cycles, generating a measured curve of molar concentration of dsDNA versus cycle number from the stored concentration values, using successive approximations, determining values of e v , e s , a, C z (n) and A z (n) which provide a best fit of the measured curve to the one or more known growth curves according to the following relation: where: C n+1 and C n are the DNA template molar concentrations at the end of the n and n + 1 cycle extension periods;

e v is in theory the template's probability of survival to the next cycle;
e s is in theory the template's probability of being synthesized;
C o is the molar concentration of starting target nucleic acid molecules;
Cp o is the starting primer molar concentration;
a is in theory the primer/complementary strand competition factor;
C z is molar concentration of the polymerase enzyme;
A z is specific activity of the polymerase;
t z is extension time in seconds;
L t is length of template in bases;
Lp is length of primers in bases; and

calculating the molar concentration of starting nucleic acid of the unknown samples by fixing e v , e s , a, C z (n), and A z (n), and adjusting the starting concentration C o to get a best fit to the measured growth curve of each unknown sample.
摘要翻译：在含有合适的缓冲液，两种互补种类的寡核苷酸引物，摩尔过量的4种核苷三磷酸酯的样品反应混合物中，在聚合酶链式反应开始时确定靶核酸分子的未知起始摩尔浓度的装置和方法， DNA聚合酶和目标核酸分子的未知起始摩尔浓度，其中以已知浓度（Cpo）提供两种引物，该方法包括添加不显着干扰反应的报道分子和在不存在双链DNA的情况下不发出强信号，但是通过反应产生的双链DNA可以每个循环至少检测一次并测量至少一次，热循环一个或多个标准反应混合物，其在各方面基本上相同，除了它们已知已知起始摩尔浓度的目标核酸分子获得一个或多个具有已知的起始摩尔浓度的靶核酸分子的生长曲线，在每个循环的至少延伸部分期间激发反应混合物，在每个循环的至少延伸部分期间检测和测量信号的强度，将所述强度至dsDNA的摩尔浓度值，并存储每个循环的每个延伸部分的摩尔浓度值，使用逐次逼近，使用逐次逼近，确定ev的值，产生dsDNA相对于循环数的摩尔浓度的测量曲线与存储的浓度值，es，a，Cz（n）和Az（n），其根据以下关系提供测量曲线与一个或多个已知生长曲线的最佳拟合：其中：Cn + 1和Cn是DNA 在n和n + 1个循环延伸期结束时的模板摩尔浓度; ev在理论上是模板生存下一个循环的概率; 理论上是模板的合成概率; Co是起始靶核酸分子的摩尔浓度; Cpo是起始底物摩尔浓度; a在理论上是引物/互补链竞争因子; Cz是聚合酶的摩尔浓度; Az是聚合酶的比活性; tz是秒的扩展时间; Lt是基数的模板长度; Lp是碱基中引物的长度; 并通过固定ev，es，a，Cz（n）和Az（n）计算未知样品的起始核酸的摩尔浓度，并调整起始浓度Co以最佳拟合每个样品的测量生长曲线未知样本

9. 发明公开

EP0059836A1 Optical beam splitter 失效
标题翻译： Optischer Strahlenteiler。
公开(公告)号：EP0059836A1
公开(公告)日：1982-09-15
申请号：EP82100648.3
申请日：1982-01-29
申请人： THE PERKIN-ELMER CORPORATION
发明人： Helms, Charles C. , Atwood, John G.
IPC分类号： G02B27/14 , G01J3/42
CPC分类号： G02B27/143 , G01J1/36
摘要： An achromatic beam splitter which is mechanically and thermally stable includes a first aperture (34), which aperture is overfilled, and an achromatic optical element (35). The aperture and element are preferably positioned in an optical plane having uniform illumination.
摘要翻译：机械和热稳定的消色差分束器包括过度填充孔径的第一孔（34）和消色差光学元件（35）。孔和元件优选地定位在具有均匀照明的光学平面中。

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