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    Automated polypeptide synthesis process
    1.
    发明授权
    Automated polypeptide synthesis process 失效
    自动多肽合成过程

    公开(公告)号：US4816513A

    公开(公告)日：1989-03-28

    申请号：US53324

    申请日：1987-05-22

    申请人： John Bridgham ,  Timothy G. Geiser ,  Michael W. Hunkapiller ,  Stephen B. H. Kent ,  Mark P. Marriott ,  Paul O. Ramstad ,  Eric S. Nordman

    发明人： John Bridgham ,  Timothy G. Geiser ,  Michael W. Hunkapiller ,  Stephen B. H. Kent ,  Mark P. Marriott ,  Paul O. Ramstad ,  Eric S. Nordman

    IPC分类号： C07K1/00 ,  B01J19/00 ,  C07K1/04 ,  C40B40/10 ,  C40B60/14 ,  C08L89/00 ,  C07C103/52

    CPC分类号： C07K1/045 ,  B01J19/0046 ,  B01J2219/00286 ,  B01J2219/00389 ,  B01J2219/00484 ,  B01J2219/00488 ,  B01J2219/0049 ,  B01J2219/00495 ,  B01J2219/005 ,  B01J2219/0059 ,  B01J2219/00596 ,  B01J2219/00689 ,  B01J2219/00695 ,  B01J2219/00725 ,  C40B40/10 ,  C40B60/14

    摘要： An apparatus is provided for automatically constructing a polypeptide of high purity, up to 50 amino acids in length, using only single couplings. The apparatus includes an activation system for receiving protected amino acids, one kind at a time, having a common vessel (an activator vessel) in which to activate each of the amino acids. Also included is a reaction vessel for containing a resin used in solid-phase peptide synthesis for attaching a peptide chain thereto. A transfer system is also provided, which operates under control of a computer, to transfer the activated species from the activation system to the reaction vessel and to transfer amino acids, reagents, gases, and solvents from one part of the apparatus to another. The activator system also includes a temperature controlled concentrator vessel in which an activator solvent is replaced by a coupling solvent to enhance the coupling of the activated species to the peptide chain in the reaction vessel. Also included in the synthesizer system is a vortexer for affecting total washing of materials in the reaction vessel and the reaction vessel itself, an automated peptide resin sampling system, and an autodelivery system for providing individual containers of amino acid to the synthesizer in the order desired in the peptide sequence. A liquid sensor system is also included to monitor transitions between gases and liquids in specific tubes in the synthesizer in order to provide input signals to the computer system for control purposes. The computer system software which controls the operation of the synthesizer is organized according to a series of menus which allows the user of the system to select individual cycles of operation for each vessel in the synthesizer. In addition, an algorithm has been developed which provides for optimum efficiency in the production of a peptide for any given selection of cycles.

    摘要翻译： 提供了一种用于仅使用单个耦合自动构建高纯度多达50个氨基酸长度的多肽的装置。 该装置包括用于一次接收受保护的氨基酸的激活系统，其具有在其中激活每个氨基酸的共同血管（活化剂容器）。 还包括用于含有固相肽合成中用于连接肽链的树脂的反应容器。 还提供了在计算机控制下操作的转移系统，以将活化物质从活化系统转移到反应容器中，并将氨基酸，试剂，气体和溶剂从设备的一部分转移到另一部分。 活化剂体系还包括温度控制的浓缩容器，其中活化剂溶剂被偶联溶剂代替以增强活化物质与反应容器中的肽链的偶联。 还包括在合成器系统中的是用于影响反应容器和反应容器本身中的材料的全部洗涤的涡流器，自动化肽树脂取样系统和用于按合适的顺序向合成器提供单个氨基酸容器的自动输送系统 在肽序列中。 还包括液体传感器系统以监测合成器中特定管中的气体和液体之间的转换，以便为控制目的向计算机系统提供输入信号。 控制合成器操作的计算机系统软件根据一系列菜单进行组织，这些菜单允许系统的用户为合成器中的每个容器选择各个操作周期。 此外，已经开发了一种算法，其提供用于任何给定选择周期的肽的生产的最佳效率。

    Automated polypeptide synthesis apparatus
    3.
    发明授权
    Automated polypeptide synthesis apparatus 失效
    自动多肽合成仪

    公开(公告)号：US5186898A

    公开(公告)日：1993-02-16

    申请号：US328488

    申请日：1989-03-24

    申请人： John Bridgham ,  Timothy G. Geiser ,  Michael W. Hunkapiller ,  Stephen B. H. Kent ,  Mark P. Marriott ,  Paul O. Ramstad ,  Eric S. Nordman

    发明人： John Bridgham ,  Timothy G. Geiser ,  Michael W. Hunkapiller ,  Stephen B. H. Kent ,  Mark P. Marriott ,  Paul O. Ramstad ,  Eric S. Nordman

    IPC分类号： B01J19/00 ,  C07K1/04 ,  C40B40/10 ,  C40B60/14

    CPC分类号： C07K1/045 ,  B01J19/0046 ,  B01J2219/00286 ,  B01J2219/00389 ,  B01J2219/00484 ,  B01J2219/00488 ,  B01J2219/0049 ,  B01J2219/00495 ,  B01J2219/005 ,  B01J2219/0059 ,  B01J2219/00596 ,  B01J2219/00689 ,  B01J2219/00695 ,  B01J2219/00725 ,  C40B40/10 ,  C40B60/14 ,  Y10S436/81

    摘要： An apparatus is provided for automatically constructing a polypeptide of high purity, up to 50 amino acids in length, using only single couplings. The apparatus includes an activation system for receiving protected amino acids, one kind at a time, having a common vessel (an activator vessel) in which to activate each of the amino acids. Also included is a reaction vessel for containing a resin used in solid-phase peptide synthesis for attaching a peptide chain thereto. A transfer system is also provided, which operates under control of a computer, to transfer the activated species from the activation system to the reaction vessel and to transfer amino acids, reagents, gases, and solvents from one part of the apparatus to another. The activator system also includes a temperature controlled concentrator vessel in which an activator solvent is replaced by a coupling solvent to enhance the coupling of the activated species to the peptide chain in the reaction vessel. Also included in the synthesizer system is a vortexer for affecting total washing of materials in the reaction vessel and the reaction vessel itself, an automated peptide resin sampling system, and an autodelivery system for providing individual containers of amino acid to the synthesizer in the order desired in the peptide sequence. A liquid sensor system is also included to monitor transitions between gases and liquids in specific tubes in the synthesizer in order to provide input signals to the computer system for control purposes. The computer system software which controls the operation of the synthesizer is organized according to a series of menus which allows the user of the system to select individual cycles of operation for each vessel in the synthesizer. In addition, an algorithm has been developed which provides for optimum efficiency in tbe production of a peptide for any given selection of cycles.

    摘要翻译： 提供了一种用于仅使用单个耦合自动构建高纯度多达50个氨基酸长度的多肽的装置。 该装置包括用于一次接收受保护的氨基酸的激活系统，其具有在其中激活每个氨基酸的共同血管（活化剂容器）。 还包括用于含有固相肽合成中用于连接肽链的树脂的反应容器。 还提供了在计算机控制下操作的转移系统，以将活化物质从活化系统转移到反应容器中，并将氨基酸，试剂，气体和溶剂从设备的一部分转移到另一部分。 活化剂体系还包括温度控制的浓缩容器，其中活化剂溶剂被偶联溶剂代替以增强活化物质与反应容器中的肽链的偶联。 还包括在合成器系统中的是用于影响反应容器和反应容器本身中的材料的全部洗涤的涡流器，自动化肽树脂取样系统和用于按合适的顺序向合成器提供单个氨基酸容器的自动输送系统 在肽序列中。 还包括液体传感器系统以监测合成器中特定管中的气体和液体之间的转换，以便为控制目的向计算机系统提供输入信号。 控制合成器操作的计算机系统软件根据一系列菜单进行组织，这些菜单允许系统的用户为合成器中的每个容器选择各个操作周期。 此外，已经开发了一种算法，其为任何给定的循环选择提供肽的生产的最佳效率。

    Restriction enzyme mediated adapter
    6.
    发明授权
    Restriction enzyme mediated adapter 有权
    限制酶介导的适配器

    公开(公告)号：US06258539B1

    公开(公告)日：2001-07-10

    申请号：US09303774

    申请日：1999-04-30

    申请人： Michael W. Hunkapiller ,  John H. Richards

    发明人： Michael W. Hunkapiller ,  John H. Richards

    IPC分类号： C12Q168

    CPC分类号： C12Q1/6837 ,  C12Q1/6809 ,  C12Q1/6874 ,  C12Q2565/501 ,  C12Q2525/191 ,  C12Q2521/313 ,  C12Q2525/179 ,  C12Q2525/161

    摘要： The present invention relate to methods and compositions for simultaneously analyzing multiple different polynucleotides of a polynucleotide composition comprising multiple diverse polynucleotide sequences. The subject methods and compositions may also be applied to analyze or identify single polynucleotides; however, the subject methods and compositions are particularly useful for analyzing large diverse populations of polynucleotides, e.g., cDNA libraries. Most embodiments of the invention involve hybridizing terminus probes (of known base sequence) to adapter-modified restriction fragment generated from polynucleotide for analysis, and subsequently joining the terminus probes and internal fragment probes to each other. The terminus probe hybridizes to bases of restriction endonuclease recognition site present at the terminus of a restriction fragment generated from the polynucleotide for analysis. The terminus probes and internal fragment probes may be marked so as to facilitate the simultaneous testing of multiple polynucleotides for the presence of many possible nucleotide base sequences. The identity or expression of a particular polynucleotide of interest may be ascertained (or at least partially determined) by producing a short identifier sequence derived from the nucleotide base sequence information obtained from (1) the hybridization of a terminus probe, and (2) the recognition site of a restriction endonuclease used to generate the polynucleotide molecule of interest. Multiple identification sequences may be obtained in parallel, thereby permitting the rapid characterization of a large number of diverse polynucleotides. Parallel processing may be achieved by differentially marking terminus probes or internal fragment probes. Parallel processing may be achieved by using ordered arrays of oligonucleotides that are terminus probes.

    摘要翻译： 本发明涉及用于同时分析包含多种不同多核苷酸序列的多核苷酸组合物的多种不同多核苷酸的方法和组合物。 主题方法和组合物也可以用于分析或鉴定单个多核苷酸; 然而，本发明的方法和组合物特别可用于分析大量不同种群的多核苷酸，例如cDNA文库。 本发明的大多数实施方案涉及将已知碱基序列的末端探针与由多核苷酸产生的衔接子修饰的限制性片段杂交用于分析，然后将末端探针和内部片段探针彼此连接。 末端探针与存在于由多核苷酸产生的限制性片段的末端存在的限制性内切核酸酶识别位点的碱基杂交，用于分析。 可以标记末端探针和内部片段探针，以促进多个多核苷酸的同时测试以存在许多可能的核苷酸碱基序列。 通过产生源自从（1）末端探针的杂交获得的核苷酸碱基序列信息的短标识符序列，可以确定（或至少部分地确定）特定感兴趣的多核苷酸的身份或表达，和（2） 用于产生感兴趣的多核苷酸分子的限制性内切核酸酶的识别位点。 可以并行获得多个鉴定序列，从而允许快速表征大量不同多核苷酸。 通过差异标记末端探针或内部片段探针可以实现并行处理。 可以通过使用作为末端探针的寡核苷酸的有序阵列来实现并行处理。

    Adapter directed expression analysis
    7.
    发明授权
    Adapter directed expression analysis 有权

    公开(公告)号：US06232067B1

    公开(公告)日：2001-05-15

    申请号：US09135381

    申请日：1998-08-17

    申请人： Michael W. Hunkapiller ,  John H. Richards

    发明人： Michael W. Hunkapiller ,  John H. Richards

    IPC分类号： C12Q168

    CPC分类号： C12Q1/6837 ,  C12Q1/6809 ,  C12Q1/6874 ,  C12Q2565/501 ,  C12Q2525/191 ,  C12Q2521/313 ,  C12Q2525/179 ,  C12Q2525/161

    摘要： The present invention relate to methods and compositions for simultaneously analyzing multiple different polynucleotides of a polynucleotide composition comprising multiple diverse polynucleotide sequences. The subject methods and compositions may also be applied to analyze or identify single polynucleotides; however, the subject methods and compositions are particularly useful for analyzing large diverse populations of polynucleotides, e.g., cDNA libraries. Most embodiments of the invention involve hybridizing terminus probes (of known base sequence) and internal fragment probes (of known base sequence) at adjacent positions on an adapter-modified restriction fragment generated from polynucleotide for analysis, and subsequently joining the terminus probes and internal fragment probes to each other. The terminus probe hybridizes to bases of restriction endonuclease recognition site present at the terminus of a restriction fragment generated from the polynucleotide for analysis. Internal fragment probes hybridizes to the same strand of the restriction fragment that the terminus probe hybridizes to and hybridizes to the restriction fragment portion of adapter-modified representative restriction fragments. The terminus probes and internal fragment probes may be marked so as to facilitate the simultaneous testing of multiple polynucleotides for the presence of many possible nucleotide base sequences. The identity or expression of a particular polynucleotide of interest may be ascertained (or at least partially determined) by producing a short identifier sequence derived from the nucleotide base sequence information obtained from (1) the hybridization of a terminus probe and an internal fragment probe, each of known base sequence, at adjacent positions on a polynucleotide of interest, and (2) the recognition site of a restriction endonuclease used to generate the polynucleotide molecule of interest. Multiple identification sequences may be obtained in parallel, thereby permitting the rapid characterization of a large number of diverse polynucleotides. Parallel processing may be achieved by differentially marking terminus probes or internal fragment probes. Parallel processing may be achieved by using ordered arrays of oligonucleotides that are terminus probes.

    Automated DNA sequencing technique
    8.
    发明授权
    Automated DNA sequencing technique 失效
    自动DNA测序技术

    公开(公告)号：US5821058A

    公开(公告)日：1998-10-13

    申请号：US361176

    申请日：1994-12-21

    申请人： Lloyd M. Smith ,  Leroy E. Hood ,  Michael W. Hunkapiller ,  Tim J. Hunkapiller ,  Charles R. Connell

    发明人： Lloyd M. Smith ,  Leroy E. Hood ,  Michael W. Hunkapiller ,  Tim J. Hunkapiller ,  Charles R. Connell

    IPC分类号： C12Q1/68 ,  G01N27/447 ,  C07H21/04 ,  C12P19/34

    CPC分类号： C12Q1/6869 ,  C12Q1/6816 ,  G01N27/44721 ,  G01N27/44726

    摘要： A process for the electrophoretic analysis of DNA fragments produced in DNA sequencing operations wherein chromophores or fluorophores are used to tag the DNA fragments produced by the sequencing chemistry and permit the detection and characterization of the fragments as they are resolved by electrophoresis through a gel. Preferably four different fragment sets are tagged with the fluorophores fluorescein, Texas Red, tetramethyl rhodamine, and 7-nitrobenzofurazan. A system for the electrophoretic analysis of DNA fragments produced in DNA sequencing operations comprising: a source of chromophore or fluorescent tagged DNA fragments; a zone for contacting an electrophoresis gel; means for introducing said tagged DNA fragments to said zone; and photometric means for monitoring said tagged DNA fragments as they move through said gel.

    摘要翻译： 用于DNA测序操作中产生的DNA片段的电泳分析的方法，其中使用发色团或荧光团标记由测序化学产生的DNA片段，并允许片段在通过凝胶电泳分离时的检测和表征。 优选地，用荧光团荧光素，德克萨斯红，四甲基罗丹明和7-硝基苯并呋咱标记四个不同的片段组。 用于在DNA测序操作中产生的DNA片段的电泳分析系统，包括：发色团或荧光标记的DNA片段; 用于接触电泳凝胶的区域; 用于将所述标记的DNA片段引入所述区域的方法; 以及用于在所述标记的DNA片段移动通过所述凝胶时监测所述标记的DNA片段的光度测量装置。

    Real time scanning electrophoresis apparatus for DNA sequencing
    10.
    发明授权
    Real time scanning electrophoresis apparatus for DNA sequencing 失效
    用于DNA测序的实时扫描电泳装置

    公开(公告)号：US4811218A

    公开(公告)日：1989-03-07

    申请号：US869421

    申请日：1986-06-02

    申请人： Michael W. Hunkapiller ,  Charles R. Connell ,  William J. Mordan ,  John D. Lytle ,  John A. Bridgham

    发明人： Michael W. Hunkapiller ,  Charles R. Connell ,  William J. Mordan ,  John D. Lytle ,  John A. Bridgham

    IPC分类号： G01N27/447 ,  G01N27/26 ,  C12Q1/68

    CPC分类号： G01N27/44721

    摘要： A real-time, automated, nucleic acid sequencing apparatus that offers high speed, definitive sequencing on many samples at the same time. The apparatus permits more than one clone to be sequenced at a time, thus vastly decreasing the time required to sequence longer fragments and reducing sequencing costs accordingly. The apparatus detects electromagnetic radiation from a plurality of lanes in an electrophoresis system wherein the plurality of lanes are arranged in a planar array. The apparatus includes an optical system for detecting the radiation at a plurality of wavelengths and is made up of a collection element, a filter for selectively transmitting the plurality of wavelengths received from the collection element, and a detection system for measuring intensity of the radiation received from the filter means. A translational stage is used for mounting the optical system and for moving the optical system parallel to the planar array in order to move the collection element back and forth across the lanes in order to receive radiation from the lanes, one lane at a time during electrophoresis. Also included is a computer system for controlling the filter and the stage, and for receiving intensity data from the detector and correlating that data with the corresponding lane and corresponding wavelengths transmitted by the filter in substantially real time.

    摘要翻译： 一种实时，自动化的核酸测序仪器，可同时为许多样品提供高速，确定的测序。 该装置允许一次克隆多于一个克隆，从而大大减少了对更长片段进行排序所需的时间，并相应降低了测序成本。 该装置在电泳系统中检测来自多个通道的电磁辐射，其中多个通道被布置成平面阵列。 该装置包括用于检测多个波长的辐射的光学系统，并且由收集元件，用于选择性地发射从收集元件接收的多个波长的滤光器以及用于测量接收的辐射强度的检测系统 从过滤装置。 平移阶段用于安装光学系统和平行于平面阵列的光学系统的移动，以便使集合元件在车道上来回移动，以便在电泳期间一次接收来自车道的辐射，一个通道 。 还包括用于控制滤波器和级的计算机系统，以及用于从检测器接收强度数据并将该数据与基本上实时地由滤波器传输的相应通道和相应波长相关联。