公开(公告)号：US06485625B1

公开(公告)日：2002-11-26

申请号：US09336848

申请日：1999-06-21

Applicant: John W. Simpson ,  Jonathan M. Rothberg ,  Gregory T. Went ,  Marie Carmen Ruiz-Martinez ,  Gregory T. Mulhern

Inventor： John W. Simpson ,  Jonathan M. Rothberg ,  Gregory T. Went ,  Marie Carmen Ruiz-Martinez ,  Gregory T. Mulhern

IPC: G01N2326

CPC classification number: B01J19/0093 ,  G01N27/44708 ,  G01N27/44721 ,  G01N27/44782 ,  G01N27/44791

Abstract: This invention is an integrated instrument for the high-capacity electrophoretic analysis of biopolymer samples. It comprises a specialized high-voltage, electrophoretic module in which the migration lanes are formed between a bottom plate and a plurality of etched grooves in a top plate, the module permitting concurrent separation of 80 or more separate samples. In thermal contact with the bottom plate is a thermal control module incorporating a plurality of Peltier heat transfer devices for the control of temperature and gradients in the electrophoretic medium. Fragments are detected by a transmission imaging spectrograph which simultaneously spatially focuses and spectrally resolves the detection region of all the migration lanes. The spectrograph comprises a transmission dispersion element and a CCD array to detect signals. Signal analysis comprises the steps of noise filtering, comparison in a configuration space with signal prototypes, and selection of the best prototype. Optionally post-processing is done by a Monte-Carlo simulated annealing algorithm to improve results. Optionally, an array of micro-reactors can be integrated into the instrument for the generation of sequencing reaction fragments directly from crude DNA samples.

Abstract translation: 本发明是用于生物聚合物样品的高容量电泳分析的综合仪器。 它包括专门的高压电泳模块，其中迁移通道形成在顶板和顶板中的多个蚀刻凹槽之间，该模块允许同时分离80个或更多个单独的样品。 与底板热接触的是热控制模块，其包含多个用于控制电泳介质中的温度和梯度的珀尔帖传热装置。 通过透射成像光谱仪检测片段，其同时空间聚焦并光谱地解析所有迁移通道的检测区域。 该光谱仪包括用于检测信号的透射色散元件和CCD阵列。 信号分析包括噪声滤波，配置空间与信号原型的比较以及最佳原型的选择。 可选地，后处理通过蒙特卡罗模拟退火算法完成以改善结果。 任选地，可以将微反应器阵列整合到仪器中，以直接从粗DNA样品产生测序反应片段。

公开(公告)号：US06193866B1

公开(公告)日：2001-02-27

申请号：US09212622

申请日：1998-12-16

Applicant: Joel S. Bader ,  Jonathan M. Rothberg ,  Michael W. Deem ,  Gregory T. Mulhern ,  Gregory T. Went ,  John Simpson ,  Steven Henck

Inventor： Joel S. Bader ,  Jonathan M. Rothberg ,  Michael W. Deem ,  Gregory T. Mulhern ,  Gregory T. Went ,  John Simpson ,  Steven Henck

IPC: G01N2726

CPC classification number: B82Y30/00 ,  G01N27/44773

Abstract: This invention relates to a method and device for separating charged particles according to their diffusivities in a separation medium by means of a spatially and temporally varying electric potential. The method is particularly suited to sizing and separating DNA fragments, to generating DNA fragment length polymorphism patterns, and to sequencing DNA through the separation of DNA sequencing reaction products. The method takes advantage of the transport of charged particles subject to an electric potential that is cycled between an off-state (in which the potential is flat) and one or more on-states, in which the potential is preferably spatially periodic with a plurality of eccentrically shaped stationary potential wells. The potential wells are at constant spatial positions in the on-state. Differences in liquid-phase diffusivities lead to charged particle separation. A preferred embodiment of the device is microfabricated. A separation medium fills physically defined separation lanes in the device. Electrodes deposited substantially transverse to the lanes create the required electric potentials. Advantageously, injection ports allow sample loading, and special gating electrodes focus the sample prior to separation. The effects of thermal gradients are minimized by placing the device in contact with a thermal control module, preferably a plurality of Peltier-effect heat transfer devices. The small size of a microfabricated device permits rapid separation in a plurality of separation lanes.

Abstract translation: 本发明涉及一种通过空间和时间上变化的电势根据其在分离介质中的扩散性分离带电粒子的方法和装置。 该方法特别适用于分选DNA片段，产生DNA片段长度多态性模式，并通过分离DNA测序反应产物对DNA进行测序。 该方法利用带电粒子的传输，该带电粒子经历在关闭状态（其中电位为平坦）和一个或多个导通状态之间循环的电位，其中电位优选为空间周期性的多个 偏心固定势阱。 势阱在导通状态下处于恒定的空间位置。 液相扩散性的差异导致带电粒子分离。 装置的优选实施例是微制造的。 分离介质填充设备中物理定义的分离通道。 基本横向于通道沉积的电极产生所需的电位。 有利地，注射端口允许样品加载，并且特殊门电极在分离之前聚焦样品。 通过将设备与热控制模块（优选多个珀耳帖效应传热装置）接触来使热梯度的影响最小化。 微型加工装置的小尺寸允许在多个分离通道中快速分离。

公开(公告)号：US5938904A

公开(公告)日：1999-08-17

申请号：US623346

申请日：1996-03-27

Applicant: Joel S. Bader ,  Jonathan M. Rothberg ,  Michael W. Deem ,  Gregory T. Mulhern ,  Gregory T. Went

Inventor： Joel S. Bader ,  Jonathan M. Rothberg ,  Michael W. Deem ,  Gregory T. Mulhern ,  Gregory T. Went

IPC: B01D57/02 ,  G01N27/26 ,  G01N27/447

CPC classification number: B82Y30/00 ,  G01N27/44773

Abstract: This invention relates to a method and device for separating charged particles according to their diffusivities in a separation medium by means of a spatially and temporarily varying electric potential. The method is particularly suited to sizing and separating DNA fragments, to generating DNA fragment length polymorphism patterns, and to sequencing DNA through the separation of DNA sequencing reaction products. The method takes advantage of the transport of charged particles subject to an electric potential that is cycled between an off-state (in which the potential is flat) and one or more on-states, in which the potential is preferably spatially periodic with a plurality of eccentrically shaped stationary potential wells. The potential wells are at constant spatial positions in the on-state. Differences in liquid-phase diffusivities lead to charged particle separation. A preferred embodiment of the device is microfabricated. A separation medium fills physically defined separation lanes in the device. Electrodes deposited substantially transverse to the lanes create the required electric potentials. Advantageously, injection ports allow sample loading, and special gating electrodes focus the sample prior to separation. The effects of thermal gradients are minimized by placing the device in contact with a thermal control module, preferably a plurality of Peltier-effect heat transfer devices. The small size of a microfabricated device permits rapid separation in a plurality of separation lanes.

Abstract translation: 本发明涉及一种通过空间和暂时变化的电位根据其在分离介质中的扩散性分离带电粒子的方法和装置。 该方法特别适用于分选DNA片段，产生DNA片段长度多态性模式，并通过分离DNA测序反应产物对DNA进行测序。 该方法利用带电粒子的传输，该带电粒子经历在关闭状态（其中电位为平坦）和一个或多个导通状态之间循环的电位，其中电位优选为空间周期性的多个 偏心固定势阱。 势阱在导通状态下处于恒定的空间位置。 液相扩散性的差异导致带电粒子分离。 装置的优选实施例是微制造的。 分离介质填充设备中物理定义的分离通道。 基本横向于通道沉积的电极产生所需的电位。 有利地，注射端口允许样品加载，并且特殊门电极在分离之前聚焦样品。 通过将设备与热控制模块（优选多个珀耳帖效应传热装置）接触来使热梯度的影响最小化。 微型加工装置的小尺寸允许在多个分离通道中快速分离。