公开(公告)号：US06455692B1

公开(公告)日：2002-09-24

申请号：US09698938

申请日：2000-10-26

申请人： Douglas T. Gjerde ,  Paul D. Taylor ,  Robert M. Haefele

发明人： Douglas T. Gjerde ,  Paul D. Taylor ,  Robert M. Haefele

IPC分类号： C07H2104

CPC分类号： C12Q1/6827 ,  C12N15/101 ,  C12Q2565/137 ,  C12Q2527/107

摘要： The present invention is directed to improved methods for detection of mutations in DNA using Denaturing Matched Ion Polynucleotide Chromatography (DMIPC). The invention includes the following aspects: analysis of PCR amplification products to identify factors that affect PCR replication fidelity; design of PCR primers; selection of an optimal temperature for performing DMIPC; selection of the mobile phase composition for gradient elution; methods for column preparation and maintenance; and methods for preparing polynucleotide samples prior to chromatographic analysis.

摘要翻译： 本发明涉及使用变性匹配离子多核苷酸色谱（DMIPC）检测DNA突变的改进方法。 本发明包括以下几个方面：分析PCR扩增产物，鉴定影响PCR复制保真度的因素; PCR引物设计; 选择用于执行DMIPC的最佳温度; 选择流动相组成进行梯度洗脱; 柱准备和维护方法; 以及在色谱分析之前制备多核苷酸样品的方法。

公开(公告)号：US06419824B1

公开(公告)日：2002-07-16

申请号：US09912568

申请日：2001-07-24

申请人： Douglas T. Gjerde ,  Christopher P. Hanna ,  Paul D. Taylor ,  Benjamin L. Legendre, Jr. ,  Robert M. Haefele

发明人： Douglas T. Gjerde ,  Christopher P. Hanna ,  Paul D. Taylor ,  Benjamin L. Legendre, Jr. ,  Robert M. Haefele

IPC分类号： B01D1508

CPC分类号： B01D15/366 ,  B01J20/287

摘要： The disclosure describes an ambient or low pressure device for separating polynucleotide fragments from a mixture of polynucleotide fragments comprises a tube having an upper solution input chamber, a lower eluant receiving chamber, and a fixed unit of separation media supported therein. The separation media has nonpolar separation surfaces which are free from multivalent cations which would react with counterion to form an insoluble polar coating on the surface of the separation media.

摘要翻译： 本公开描述了用于从多核苷酸片段的混合物分离多核苷酸片段的环境或低压装置，包括具有上溶液输入室，下洗脱液接收室和支撑在其中的分离介质的固定单元的管。 分离介质具有非极性分离表面，其不含多价阳离子，其将与抗衡离子反应以在分离介质的表面上形成不溶性极性涂层。

公开(公告)号：US06265168B1

公开(公告)日：2001-07-24

申请号：US09318407

申请日：1999-05-25

申请人： Douglas T. Gjerde ,  Christopher P. Hanna ,  Paul D. Taylor ,  Benjamin L. Legendre, Jr. ,  Robert M. Haefele

发明人： Douglas T. Gjerde ,  Christopher P. Hanna ,  Paul D. Taylor ,  Benjamin L. Legendre, Jr. ,  Robert M. Haefele

IPC分类号： C12Q168

CPC分类号： B01D15/366 ,  B01J20/287

摘要： A method for removing a target DNA fragment having a predetermined base-pair length from a mixture of DNA fragments comprises the following steps. A mixture of DNA fragments which may contain the target DNA fragments is applied to a separation column containing media having a nonpolar, nonporous surface, the mixture of DNA fragments being in a first solvent mixture containing a counterion and a DNA binding concentration of driving solvent in a cosolvent. The target DNA fragments are separated from the media by contacting it with a second solvent solution containing a counterion and a concentration of driving solvent in cosolvent which has been predetermined to remove DNA fragments having the target DNA fragment base pair length from the media. The target DNA fragments can be collected and optionally amplified. When the method is being applied to collect a putative fragment, if present, no DNA fragments having the base pair length of the target DNA could be present in the mixture. Alternatively, DNA fragments having the base pair length of the target DNA are present in the mixture. The disclosure also describes an ambient or low pressure device for separating polynucleotide fragments from a mixture of polynucleotide fragments comprises a tube having an upper solution input chamber, a lower eluant receiving chamber, and a fixed unit of separation media supported therein. The separation media has nonpolar separation surfaces which are free from multivalent cations which would react with counterion to form an insoluble polar coating on the surface of the separation media.

摘要翻译： 从DNA片段的混合物中除去具有预定碱基对长度的靶DNA片段的方法包括以下步骤。 将含有靶DNA片段的DNA片段的混合物应用于含有非极性，无孔表面的培养基的分离塔，DNA片段的混合物在含有抗衡离子的第一溶剂混合物和驱动溶剂的DNA结合浓度 共溶剂 通过使目标DNA片段与含有抗衡离子的溶剂溶液和在助溶剂中的驱动溶剂浓度的第二溶剂溶液接触来分离，所述助溶剂已被预先从介质中除去具有靶DNA片段碱基对长度的DNA片段。 可以收集目标DNA片段并任选地扩增。 当该方法被应用于收集假定片段时（如果存在），则在混合物中不能存在具有目标DNA碱基对长度的DNA片段。 或者，具有靶DNA的碱基对长度的DNA片段存在于混合物中。 本公开还描述了用于从多核苷酸片段的混合物分离多核苷酸片段的环境或低压装置，包括具有上溶液输入室，下洗脱液接收室和支撑在其中的分离介质的固定单元的管。 分离介质具有非极性分离表面，其不含多价阳离子，其将与抗衡离子反应以在分离介质的表面上形成不溶性极性涂层。

公开(公告)号：US06210885B1

公开(公告)日：2001-04-03

申请号：US09169440

申请日：1998-10-09

申请人： Douglas T. Gjerde ,  Paul D. Taylor ,  Robert M. Haefele

发明人： Douglas T. Gjerde ,  Paul D. Taylor ,  Robert M. Haefele

IPC分类号： C12Q168

CPC分类号： C12Q1/6816 ,  C12Q1/6827 ,  Y10T436/143333 ,  C12Q2565/137 ,  C12Q2527/107

摘要： Covalently bound non-polar tags are used to increase the retention times of double stranded polynucleotides on Matched Ion Polynucleotide Chromatography (MIPC) columns. In doing so, separations of DNA mixture components is improved. Additionally, when the non-polar tags are fluorophores, detection limits are also greatly reduced. Strategically tagged primers are used in conduction with PCR to produce DNA fragments having specifically tagged strands. This improves mutation detection by MIPC in several ways. Separations are improved, detection sensitivity is enhanced, and non-stoichiometric addition of wild type DNA prior to hybridization is now possible since only tagged fragments will be observed with a fluorescence detector. Non-polar tags are also used as a novel alternative to G-C clamping during MIPC under partially denaturing conditions. Reversible DNA binding dyes, such as DNA intercalator dyes and DNA groove binding dyes, are used to reduce the detection limit of polynucleotides separated by MIPC.

摘要翻译： 共价结合的非极性标签用于增加匹配离子多核苷酸色谱（MIPC）色谱柱上双链多核苷酸的保留时间。 在这样做时，提高了DNA混合物组分的分离。 此外，当非极性标签是荧光团时，检测限也大大降低。 策略性标记的引物用于PCR导入以产生具有特异性标记的链的DNA片段。 这可以通过多种方式改善MIPC的突变检测。 分离改善，检测灵敏度提高，杂交前非化学计量添加野生型DNA是可能的，因为只有标记的片段才能用荧光检测器观察到。 非极性标签也被用作MIPC在部分变性条件下G-C夹持的新型替代物。 使用可逆DNA结合染料，例如DNA嵌入剂染料和DNA沟槽结合染料，以降低由MIPC分离的多核苷酸的检测限。

公开(公告)号：US6103112A

公开(公告)日：2000-08-15

申请号：US295474

申请日：1999-04-19

申请人： John E. Sutton ,  Douglas T. Gjerde ,  Paul D. Taylor

发明人： John E. Sutton ,  Douglas T. Gjerde ,  Paul D. Taylor

IPC分类号： B01D15/08 ,  B01D15/12 ,  B01D15/16 ,  B01D15/20 ,  B01D15/36 ,  B01D15/38 ,  B01J45/00 ,  C12N15/10 ,  C12Q1/68 ,  G01N30/14 ,  G01N30/30 ,  G01N30/52 ,  G01N30/54

CPC分类号： B01D15/125 ,  B01D15/161 ,  B01D15/20 ,  B01D15/366 ,  B01D15/38 ,  B01J45/00 ,  C12N15/101 ,  C12Q1/6827 ,  C12Q1/6834 ,  G01N30/30 ,  G01N30/54 ,  G01N2030/528 ,  G01N30/14

摘要： A liquid chromatography apparatus with stationary and mobile phase temperature controls suitable for polynucleotide separations by MIPC and DMIPC processes. The apparatus includes heater means with a temperature control system; a matched ion polynucleotide chromatography separation column having an inlet end; a coil of capillary tubing having an inlet end and an outlet end. The outlet end of the capillary tubing is connected with the inlet end of the separation column. The inlet end of the capillary tubing comprising means for receiving process liquid, the tubing having a length of from 6 to 400 cm having a linear tubing length of heating means. The separation column and the coil of capillary tubing are enclosed in the heater means. The capillary tubing preferably is PEEK or titanium. The heater means can be an air batch oven. Preferably, it is a heat-conducting block having a first heat transfer surface, a separation column receptacle, and a capillary coil receptacle. A separation column is positioned within the separation column receptacle in heat conducting relationship with an inner wall thereof. A coil of capillary tubing is positioned in the capillary coil receptacle, the outer extremities of the coil being in heat conducting relationship with an inner wall of the capillary coil receptacle. Optimally, the heating means is a Peltier heating and cooling unit in heat conducting relationship with a heat transfer surface of the heating block.

公开(公告)号：US6027898A

公开(公告)日：2000-02-22

申请号：US136084

申请日：1998-08-18

申请人： Douglas T. Gjerde ,  Paul D. Taylor

发明人： Douglas T. Gjerde ,  Paul D. Taylor

IPC分类号： C12Q1/68

CPC分类号： C12Q1/68

摘要： A method for analyzing a sample of double stranded DNA to determine the presence of a mutation therein comprises contacting the sample with a mutation site binding reagent, and chromatographically separating and detecting the product. The chromatographic separation can be performed using Matched Ion Polynucleotide Chromatography, size exclusion chromatography, ion exchange chromatography, or reverse phase chromatography. The mutation site binding reagent can be an enzyme or a non-proteinaceous chemical reagent. In one embodiment, a mutation site binding reagent binds to the site of mutation and alters the chromatographic retention time. In another embodiment, a mutation site binding reagent cleaves at the site of mutation, resulting in an increase in the number of fragments.

摘要翻译： 用于分析双链DNA样品以确定其中突变的存在的方法包括使样品与突变位点结合试剂接触，并色谱分离和检测产物。 色谱分离可以使用匹配离子多核苷酸色谱，大小排阻层析，离子交换层析或反相色谱法进行。 突变位点结合试剂可以是酶或非蛋白质化学试剂。 在一个实施方案中，突变位点结合试剂结合突变位点并改变色谱保留时间。 在另一个实施方案中，突变位点结合试剂在突变位点切割，导致片段数量的增加。

公开(公告)号：US5319041A

公开(公告)日：1994-06-07

申请号：US38723

申请日：1993-03-26

申请人： Yuanzhen Zhong ,  Hemant Parikh ,  Paul D. Taylor ,  Terry E. Smith

发明人： Yuanzhen Zhong ,  Hemant Parikh ,  Paul D. Taylor ,  Terry E. Smith

IPC分类号： C08F2/00 ,  C08F226/10 ,  C08F2/06 ,  C08F4/04 ,  C08F4/34 ,  C08F218/08

CPC分类号： C08F226/10

摘要： This invention relates to an improved process for producing a clear solution of vinylpyrrolidone (VP) and vinyl acetate (VA) monomers which comprises, in an anhydrous system,(a) reacting a C.sub.2 to C.sub.3 alcoholic solution containing said VA monomer and between about 60 and about 80 wt. % portion of total VP monomer in the presence of between about 0.05 and about 0.3 wt. % of a free radical initiator selected from the group consisting of tertamylperoxy pivalate and 2,2-azobis(2-methylbutyronitrile) or a mixture thereof for a period of from about 5 to about 10 hours, at a temperature between about 60.degree. and about 110.degree. C.;(b) gradually adding the remaining VP monomer in C.sub.2 to C.sub.3 alcohol solution at reaction temperature after the addition of the VA monomer is complete;(c) raising the temperature of the resulting reaction mixture to between about 110.degree. and about 150.degree. C.;(d) continuing the polymerization reaction at said higher temperature in the presence of from about 0.11 to about 0.6 wt. %, based on total monomers, of 2,5-dimethyl-2,5-bis(tert-butylperoxy) hexane and(e) cooling the resulting reaction mixture and recovering a homogeneous VP/VA copolymer C.sub.2 to C.sub.3 alcohol solution containing less than 100 ppm of residual vinylpyrrolidone monomer or vinyl acetate residual monomer.

摘要翻译： 本发明涉及一种用于生产乙烯基吡咯烷酮（VP）和乙酸乙烯酯（VA）单体的透明溶液的改进方法，其包括在无水体系中，（a）使含有所述VA单体的C2至C3醇溶液与约60 和约80wt。 在约0.05至约0.3重量％存在下的总VP单体的％部分。 ％的过氧化新戊酸叔丁酯和2,2-偶氮双（2-甲基丁腈）的自由基引发剂或其混合物在约5至约10小时之间，温度在约60°至约 110℃。 （b）在加入VA单体完成后，在反应温度下逐渐加入剩余的VP单体在C2〜C3醇溶液中; （c）将所得反应混合物的温度提高至约110℃至约150℃; （d）在约0.11至约0.6重量％的存在下，在所述较高温度下继续进行聚合反应。 ％，基于总单体，2,5-二甲基-2,5-双（叔丁基过氧））己烷和（e）冷却所得反应混合物，并回收均匀的VP / VA共聚物C2至C3醇溶液含有少于 100ppm的残余乙烯基吡咯烷酮单体或乙酸乙烯酯残留单体。

公开(公告)号：US5183946A

公开(公告)日：1993-02-02

申请号：US852421

申请日：1992-03-16

申请人： Kou-Chang Liu ,  Paul D. Taylor

发明人： Kou-Chang Liu ,  Paul D. Taylor

IPC分类号： C07C41/08 ,  C07C43/178

CPC分类号： C07C41/08 ,  C07C2101/14 ,  C07C2101/18

摘要： The invention relates to a vinyloxy hydroxyalkyl cycloalkylene having the formulaHO(CH.sub.2).sub.x --A--(CH.sub.2).sub.x, OCH.dbd.CH.sub.2wherein x has a value of from 1 to 10; x' has a value of from 0 to 10 and A is a cycloalkylene group having from 3 to 8 carbon atoms in the ring and is optionally substituted with lower alkyl. The invention also relates to the method for synthesizing the above compounds.

摘要翻译： 本发明涉及具有式HO（CH 2）x -A-（CH 2）x，OCH = CH 2的乙烯基氧基羟基烷基亚环烷基，其中x具有1至10的值; x'具有0至10的值，并且A是在环中具有3至8个碳原子的亚环烷基，并且任选被低级烷基取代。 本发明还涉及上述化合物的合成方法。

公开(公告)号：US4954582A

公开(公告)日：1990-09-04

申请号：US404468

申请日：1989-09-08

申请人： Kou-Chang Liu ,  Paul D. Taylor

发明人： Kou-Chang Liu ,  Paul D. Taylor

IPC分类号： C07C43/23 ,  C08F16/32

CPC分类号： C08F16/32 ,  C07C43/23

摘要： The invention relates to benzyloxy vinyl ethers having the structure ##STR1## wherein R.sub.1 is lower alkyl;p is an integer having a value of from 0 to 2;q is an integer having a value of from 1 to 20;r is an integer having a value of from 1 to 6;n is an integer having a value of from 0 to 20;m is an integer having a value of from 0 to 1;Y is sulfur or oxygen;X is hydrogen or methyl andR and R.sub.2 are each independently a radical having from 2 to 20 carbon atoms and are selected from the group of alkylene, arylene, aralkenylene and alkarylene optionally substituted with lower alkyl.The invention also relates to the process for preparing the above multifunctional benzyloxy vinyl ethers.

摘要翻译： 本发明涉及具有以下结构的苄氧基乙烯基醚：其中R 1为低级烷基; p是0至2的整数; q是1至20的整数; r为1〜6的整数， n是0至20的整数; m是0至1的整数; Y是硫或氧; X是氢或甲基，R和R 2各自独立地是具有2至20个碳原子的基团，并且选自任选被低级烷基取代的亚烷基，亚芳基，亚链烯基和亚烷芳基。 本发明还涉及制备上述多官能苄氧基乙烯基醚的方法。

公开(公告)号：US4797382A

公开(公告)日：1989-01-10

申请号：US126065

申请日：1987-11-27

申请人： Waldo De Thomas ,  Paul D. Taylor

发明人： Waldo De Thomas ,  Paul D. Taylor

IPC分类号： B01J23/89 ,  C07C27/00 ,  C07C29/10 ,  C07C29/145 ,  C07C29/149 ,  C07C31/20 ,  C07C67/00 ,  B01J23/58 ,  B01J23/64 ,  B01J23/72

CPC分类号： C07C29/149 ,  B01J23/8926 ,  B01J23/8946

摘要： The invention relates to a novel supported catalyst comprising (a) copper, (b) a metal having an atomic weight greater than 100 and selected from Groups VIIB and VIII of the Periodic Table and (c) an alkali metal in a weight ratio of about 8-40:0.05-5:1.5-10 and to the use of the catalyst in the hydrogenation of butyrolactone to 1,4-butanediol.