公开(公告)号：US5928907A

公开(公告)日：1999-07-27

申请号：US752973

申请日：1996-12-02

申请人： Timothy M. Woudenberg ,  Kevin S. Bodner ,  Charles R. Connell ,  Alan M. Ganz ,  Lincoln J. McBride ,  Paul G. Saviano ,  John Shigeura ,  David H. Tracy ,  Eugene F. Young ,  Linda G. Lee

发明人： Timothy M. Woudenberg ,  Kevin S. Bodner ,  Charles R. Connell ,  Alan M. Ganz ,  Lincoln J. McBride ,  Paul G. Saviano ,  John Shigeura ,  David H. Tracy ,  Eugene F. Young ,  Linda G. Lee

IPC分类号： C12N15/09 ,  C12P19/34 ,  C12Q1/68 ,  G01N21/03 ,  G01N21/64 ,  G01N21/75 ,  G01N21/78 ,  G01N33/50 ,  G01N33/58

CPC分类号： G01N21/6428 ,  C12Q1/6818 ,  C12Q1/6851 ,  C12Q1/686 ,  G01N21/0303 ,  G01N2021/6484

摘要： 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.

摘要翻译： 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，反应混合物含有（i）第一荧光指示剂，其能够产生第一荧光信号，其强度与扩增产物的量成比例 在由激发束照射的反应混合物的体积中和（ii）均匀分布在整个反应混合物中的第二荧光指示器，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：US5827480A

公开(公告)日：1998-10-27

申请号：US823735

申请日：1997-03-25

申请人： Lawrence A. Haff ,  Enrico Picozza ,  Will Bloch ,  Timothy M. Woudenberg

发明人： Lawrence A. Haff ,  Enrico Picozza ,  Will Bloch ,  Timothy M. Woudenberg

IPC分类号： C12N15/09 ,  B01J19/00 ,  B01L7/00 ,  C12M1/00 ,  C12Q1/68 ,  C40B40/06 ,  C40B60/14 ,  G01N35/00 ,  G01N21/01 ,  C12M1/02 ,  G01N21/62 ,  G01N33/50

CPC分类号： B01L7/525 ,  B01J19/0046 ,  B01L7/52 ,  B01L9/065 ,  B01J2219/00495 ,  B01J2219/00722 ,  B01L2300/1883 ,  C12Q1/686 ,  C40B40/06 ,  C40B60/14 ,  G01N2035/00237 ,  Y10T436/116664

摘要： Apparatus and method for performing a nucleic acid amplification reaction and preferably a polymerase chain reaction (PCR) in a reaction mixture in at least one capillary tube. Several different embodiments are disclosed. One embodiment cycles a sample through a capillary tube loop passing through two thermostatted fluid baths. Another embodiment has the capillary tube routed alternatingly between two heat exchangers to that the sample makes only one pass through the tube. Other embodiments maintain the heat exchangers stationary and translate the samples between them. Still further embodiments maintain the samples stationary and either automatically translate or rotate the heat exchangers past the samples contained within the capillary tubes to perform the thermal cycles necessary for the amplification reaction.

摘要翻译： 用于在至少一个毛细管中的反应混合物中进行核酸扩增反应和优选聚合酶链式反应（PCR）的装置和方法。 公开了几个不同的实施例。 一个实施例通过穿过两个恒温流体浴的毛细管回路循环样品。 另一个实施例中，毛细管在两个热交换器之间交替布置，使得样品仅穿过管中一次。 其他实施例保持热交换器平稳，并在它们之间平移样品。 另外的实施例保持样品静止，并且自动地将热交换器平移或旋转通过毛细管内包含的样品，以进行扩增反应所需的热循环。

公开(公告)号：US5602756A

公开(公告)日：1997-02-11

申请号：US569606

申请日：1995-12-08

申请人： John G. Atwood ,  Albert C. Mossa ,  Lisa M. Goven ,  Fenton Williams ,  Timothy M. Woudenberg ,  Marcel Margulies ,  Robert P. Ragusa ,  Richard Leath ,  Clive Miles

发明人： John G. Atwood ,  Albert C. Mossa ,  Lisa M. Goven ,  Fenton Williams ,  Timothy M. Woudenberg ,  Marcel Margulies ,  Robert P. Ragusa ,  Richard Leath ,  Clive Miles

IPC分类号： C12M1/00 ,  B01L3/00 ,  B01L3/14 ,  B01L7/00 ,  B01L9/00 ,  C12M1/02 ,  C12M1/34 ,  C12M1/36 ,  C12M1/38 ,  C12M1/40 ,  C12N15/00 ,  C12N15/09 ,  C12P19/34 ,  C12Q20060101 ,  C12Q1/00 ,  C12Q1/24 ,  C12Q1/68 ,  G01N35/00 ,  G05B13/00 ,  G05B15/00 ,  G05D23/00 ,  G05D23/19 ,  G06G7/58

CPC分类号： G05D23/1935 ,  B01L3/50851 ,  B01L3/50853 ,  B01L7/00 ,  B01L7/52 ,  G05D23/1917 ,  G05D23/1919 ,  B01L2200/147 ,  B01L2300/042 ,  B01L2300/046 ,  B01L2300/0829 ,  B01L2300/1822 ,  B01L2300/1827 ,  B01L2300/185 ,  B01L3/5082

摘要： An instrument for performing highly accurate PCR employing a sample block in microtiter tray format. The sample block has local balance and local symmetry. A three zone film heater controlled by a computer and ramp cooling solenoid valves also controlled by the computer for gating coolant flow through the block controls the block temperature. Constant bias cooling is used for small changes. Sample temperature is calculated instead of measured. A platen deforms plastic caps to apply a minimum acceptable threshold force for seating the tubes and thermally isolates them. A cover isolates the block. The control software includes diagnostics. An install program tests and characterizes the instrument. A new user interface is used. Disposable, multipiece plastic microtiter trays to give individual freedom to sample tubes are taught.

摘要翻译： 使用微量滴定盘格式的样品块进行高精度PCR的仪器。 样品块具有局部平衡和局部对称性。 由计算机控制的三区薄膜加热器也由计算机控制的斜坡冷却电磁阀用于选通通过块的冷却剂流控制块温度。 恒定偏压冷却用于小的变化。 计算样品温度而不是测量。 压板使塑料盖变形，以施加最小可接受的阈值力，用于安置管并将其热隔离。 盖子隔离块。 控制软件包括诊断。 安装程序测试和表征仪器。 使用新的用户界面。 教授了一次性的多片塑料微量滴定塔，以给予样品管的个体自由度。