公开(公告)号：US06569979B1

公开(公告)日：2003-05-27

申请号：US09659442

申请日：2000-09-08

申请人： Todd C. Strother ,  Lloyd M. Smith ,  Robert J. Hamers

发明人： Todd C. Strother ,  Lloyd M. Smith ,  Robert J. Hamers

IPC分类号： C08G7700

CPC分类号： B82Y30/00 ,  B05D1/185 ,  B82Y10/00 ,  B82Y40/00 ,  H01L51/0093

摘要： Chemically-modified surfaces on unoxidized carbon, silicon, and germanium substrates are disclosed. Ultraviolet radiation mediates the reaction of protected &ohgr;-modified, &agr;-unsaturated aminoalkenes (preferred) with hydrogen-terminated carbon, silicon, or germanium surfaces. Removal of the protecting group yields an aminoalkane-modified silicon surface. These amino groups can be coupled to terminal-modified oligonucleotides using a bifunctional crosslinker, thereby permitting the preparation of modified surfaces and arrays. Methods for controlling the surface density of molecules attached to the substrate are also disclosed.

摘要翻译： 公开了未氧化的碳，硅和锗衬底上的化学修饰的表面。 紫外线辐射介导受保护的ω-改性的α-不饱和氨基烯烃（优选）与氢封端的碳，硅或锗表面的反应。 去除保护基产生氨基烷烃改性的硅表面。 这些氨基可以使用双功能交联剂与末端修饰的寡核苷酸偶联，从而允许制备改性的表面和阵列。 还公开了用于控制附着于基底的分子的表面密度的方法。

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。

公开(公告)号：US5747249A

公开(公告)日：1998-05-05

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 ,  C12Q2563/107 ,  C12Q2535/101

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-nitro-benzofurazan. 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.

公开(公告)号：US5591841A

公开(公告)日：1997-01-07

申请号：US317102

申请日：1994-09-30

申请人： Huamin Ji ,  Lloyd M. Smith

发明人： Huamin Ji ,  Lloyd M. Smith

IPC分类号： C12N15/10 ,  C12Q1/68 ,  C07H21/04

CPC分类号： C12Q1/6839 ,  C12N15/1003

摘要： A single-step capture of a target supercoiled double-stranded DNA molecule is accomplished by forming a local triple-helix among two strands of the supercoiled circular DNA and an oligonucleotide probe. The oligonucleotide is bound to an immobilizing support which facilitates the immobilization and purification of target DNA molecules. Non-target DNA molecules and other contaminating cellular material are easily removed by washing. The triple-helical structure is destabilized by raising the pH, leaving purified target DNA in the supernatant and reusable affinity capture oligonucleotide secured to the immobilizing support.

摘要翻译： 目标超螺旋双链DNA分子的单步捕获通过在超螺旋环状DNA的两条链和寡核苷酸探针中形成局部三螺旋来实现。 寡核苷酸与固定化载体结合，有利于靶DNA分子的固定和纯化。 非目标DNA分子和其他污染细胞材料容易通过洗涤去除。 三螺旋结构通过提高pH而不稳定，将纯化的靶DNA留在上清液中，并将可重复使用的亲和捕获寡核苷酸固定在固定支持物上。

公开(公告)号：US5376549A

公开(公告)日：1994-12-27

申请号：US14944

申请日：1993-02-05

申请人： Richard A. Guilfoyle ,  Lloyd M. Smith

发明人： Richard A. Guilfoyle ,  Lloyd M. Smith

IPC分类号： C12N15/70

CPC分类号： C12N15/70

摘要： A vector comprising a filamentous phage sequence containing a first copy of filamentous phage gene X and other sequences necessary for the phage to propagate is disclosed. The vector also contains a second copy of filamentous phage gene X downstream from a promoter capable of promoting transcription in a bacterial host. In a preferred form of the present invention, the filamentous phage is M13 and the vector additionally includes a restriction endonuclease site located in such a manner as to substantially inactivate the second gene X when a DNA sequence is inserted into the restriction site.

摘要翻译： 公开了一种载体，其包含含有丝状噬菌体基因X的第一拷贝的丝状噬菌体序列和噬菌体传播所必需的其它序列。 载体还含有能够在细菌宿主中促进转录的启动子下游的丝状噬菌体基因X的第二拷贝。 在本发明的优选形式中，丝状噬菌体是M13，并且所述载体另外包含限制性内切核酸酶位点，所述限制性内切核酸酶位点以当将DNA序列插入限制性位点时基本上灭活第二基因X.

公开(公告)号：US5118802A

公开(公告)日：1992-06-02

申请号：US661913

申请日：1991-02-27

申请人： Lloyd M. Smith ,  Steven Fung ,  Robert J. Kaiser, Jr.

发明人： Lloyd M. Smith ,  Steven Fung ,  Robert J. Kaiser, Jr.

IPC分类号： C07H21/00 ,  C12Q1/68

CPC分类号： C07H21/00 ,  C12Q1/6816

摘要： The invention consists of compounds and methods for the synthesis of oligonucleotides which contain one or more free aliphatic amino groups attached to the sugar moieties of the nucleoside subunits. The synthetic method is versatile and general, permitting amino groups to be selectively placed at any position on oligonucleotides of any composition or length which is attainable by current DNA synthetic methods. Fluorescent dyes or other detectable moieties may be covalently attached to the amino groups to yield the corresponding modified oligonucleotide.

摘要翻译： 本发明由用于合成寡核苷酸的化合物和方法组成，其含有连接至核苷亚基的糖部分的一个或多个游离脂族氨基。 合成方法是通用且通用的，允许将氨基选择性地置于任何可通过当前DNA合成方法获得的组合物或长度的寡核苷酸上的任何位置。 荧光染料或其它可检测部分可以共价连接到氨基上以产生相应的修饰的寡核苷酸。

公开(公告)号：US5015733A

公开(公告)日：1991-05-14

申请号：US287387

申请日：1988-12-19

申请人： Lloyd M. Smith ,  Steven Fund ,  Robert J. Kaiser, Jr.

发明人： Lloyd M. Smith ,  Steven Fund ,  Robert J. Kaiser, Jr.

IPC分类号： C07H21/00 ,  C12Q1/68

CPC分类号： C07H21/00 ,  C12Q1/6816

摘要： The invention consists of compounds and methods for the synthesis of oligonucleotides which contain one or more free aliphatic amino groups attached to the sugar moieties of the nucleoside subunits. The synthetic method is versatile and general, permitting amino groups to be selectively placed at any position on oligonucleotides of any composition or length which is attainable by current DNA synthetic methods. Fluorescent dyes or other detectable moieites may be covalently attached to the amino groups to yield the corresponding modified oligonucleotide.

摘要翻译： 本发明由用于合成寡核苷酸的化合物和方法组成，其含有连接至核苷亚基的糖部分的一个或多个游离脂族氨基。 合成方法是通用且通用的，允许将氨基选择性地置于任何可通过当前DNA合成方法获得的组合物或长度的寡核苷酸上的任何位置。 可以将荧光染料或其它可检测的硅氧烷共价连接到氨基上，得到相应的修饰寡核苷酸。

公开(公告)号：US08686375B2

公开(公告)日：2014-04-01

申请号：US13625731

申请日：2012-09-24

申请人： Robert H. Blick ,  Lloyd M. Smith ,  Michael Westphall ,  Hua Qin

发明人： Robert H. Blick ,  Lloyd M. Smith ,  Michael Westphall ,  Hua Qin

IPC分类号： G01T1/00

CPC分类号： G01N23/00 ,  H01J47/001 ,  H01J49/025

摘要： An active detector and methods for detecting molecules, including large molecules such as proteins and oligonucleotides, at or near room temperature based on the generation of electrons via field emission (FE) and/or secondary electron emission (SEE). The detector comprises a semiconductor membrane having an external surface that is contacted by one or more molecules, and an internal surface having a thin metallic layer or other type of electron emitting layer. The kinetic energy of molecules contacting the semiconductor membrane is transferred through the membrane and induces the emission of electrons from the emitting layer. An electron detector, which optionally includes means for electron amplification, is positioned to detect the emitted electrons.

摘要翻译： 基于通过场发射（FE）和/或二次电子发射（SEE）产生电子，在室温或室温下检测分子（包括大分子如蛋白质和寡核苷酸）的主动检测器和方法。 检测器包括具有与一个或多个分子接触的外表面的半导体膜和具有薄金属层或其它类型的电子发射层的内表面。 接触半导体膜的分子的动能通过膜转移并诱导从发射层发射电子。 可选地包括用于电子放大的装置的电子检测器被定位成检测发射的电子。

公开(公告)号：US08592216B2

公开(公告)日：2013-11-26

申请号：US12759950

申请日：2010-04-14

申请人： Brian L. Frey ,  April L. Jue ,  Casey J. Krusemark ,  Lloyd M. Smith ,  Joshua J. Coon

发明人： Brian L. Frey ,  April L. Jue ,  Casey J. Krusemark ,  Lloyd M. Smith ,  Joshua J. Coon

IPC分类号： G01N37/00

CPC分类号： C07K1/13

摘要： The present invention provides methods for enhancing the fragmentation of peptides for mass spectrometry by modifying the peptides with a tagging reagent containing a functional group, such as a tertiary amine, having a greater gas-phase basicity than the amide backbone of the peptide. These high gas-phase basicity functional groups are attached to a peptide by reacting the tagging reagent to one or more available carboxylic acid groups of the peptide. Linking these high gas-phase functional groups to the peptides leads to higher charge state ions from electrospray ionization mass spectrometry (ESI-MS), which fragment more extensively during fragmentation techniques, particularly non-ergodic fragmentation techniques such as electron capture dissociation (ECD) and electron transfer dissociation (ETD).

摘要翻译： 本发明提供了通过用含有与肽的酰胺主链相比具有更大的气相碱度的官能团的标记试剂（例如叔胺）修饰肽来增强肽质谱分离的方法。 这些高气相碱性官能团通过使标记试剂与肽的一个或多个可用的羧酸基团反应而连接到肽上。 将这些高气相官能团连接到肽导致来自电喷雾离子化质谱（ESI-MS）的更高电荷状态离子，其在片段化技术期间更广泛地进行片段，特别是非遍历断裂技术，例如电子俘获解离（ECD） 和电子转移离解（ETD）。

公开(公告)号：US20100330680A1

公开(公告)日：2010-12-30

申请号：US12759950

申请日：2010-04-14

申请人： Brian L. Frey ,  April L. Jue ,  Casey J. Krusemark ,  Lloyd M. Smith ,  Joshua J. Coon

发明人： Brian L. Frey ,  April L. Jue ,  Casey J. Krusemark ,  Lloyd M. Smith ,  Joshua J. Coon

IPC分类号： G01N37/00 ,  C07K1/13 ,  C07K1/14 ,  C07K2/00

CPC分类号： C07K1/13

摘要： The present invention provides methods for enhancing the fragmentation of peptides for mass spectrometry by modifying the peptides with a tagging reagent containing a functional group, such as a tertiary amine, having a greater gas-phase basicity than the amide backbone of the peptide. These high gas-phase basicity functional groups are attached to a peptide by reacting the tagging reagent to one or more available carboxylic acid groups of the peptide. Linking these high gas-phase functional groups to the peptides leads to higher charge state ions from electrospray ionization mass spectrometry (ESI-MS), which fragment more extensively during fragmentation techniques, particularly non-ergodic fragmentation techniques such as electron capture dissociation (ECD) and electron transfer dissociation (ETD).

摘要翻译： 本发明提供了通过用含有与肽的酰胺主链相比具有更大的气相碱度的官能团的标记试剂（例如叔胺）修饰肽来增强肽质谱分离的方法。 这些高气相碱性官能团通过使标记试剂与肽的一个或多个可用的羧酸基团反应而连接到肽上。 将这些高气相官能团连接到肽导致来自电喷雾离子化质谱（ESI-MS）的更高电荷状态离子，其在片段化技术期间更广泛地进行片段，特别是非遍历断裂技术，例如电子俘获解离（ECD） 和电子转移离解（ETD）。