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公开(公告)号：US06136195A
公开(公告)日：2000-10-24
申请号：US285331
申请日：1999-04-02
IPC分类号： B01D15/08
CPC分类号： B01D15/366 , B01D15/203 , G01N30/50 , G01N2030/201 , G01N2030/202
摘要： An apparatus for effecting base pair length separations of DNA fragments by matched ion paired chromatography comprising a separation column containing separation media having non-polar DNA separation surfaces, separation solution supply means, and a separation solution conduit communicating with the separation column and the separation solution supply means, and a cleaning solution valve means positioned in the separation solution conduit for injecting cleaning solution into the separation solution conduit. A process for cleaning the non-polar DNA separation surfaces in the apparatus comprising interrupting the flow of separation solvent with a block of cleaning solution injected into the flow of separation solution passing to the column, the cleaning solution containing agent which removes accumulated residues from the non-polar surface. The cleaning solution can have an alkaline pH and contain a chelating agent such as EDTA.
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公开(公告)号：US06355791B1
公开(公告)日：2002-03-12
申请号：US09562069
申请日：2000-05-01
IPC分类号： C07H2104
CPC分类号： B01D15/366 , B01D15/20 , B01J20/285 , C08F8/02 , C08F8/30 , C08F212/08 , C08F230/08 , C12N15/101
摘要： Non-polar polymeric separation media, such as beads or monoliths, are suitable for chromatographic separation of mixtures of polynucleotides when the surface of the media are unsubstituted or substituted with a hydrocarbon group having from one to 1,000,000 carbons and when the surfaces are substantially free from multivalent cation contamination. The polymeric media provide efficient separation of polynucleotides using Matched ion Polynucleotide Chromatography. Methods for maintaining and storing the polymeric media include treatment with multivalent cation binding agents.
摘要翻译： 非极性聚合物分离介质，例如珠粒或整料，适用于当介质的表面未被取代或被具有1至1,000,000个碳原子的烃基取代时，多个核苷酸混合物的色谱分离，并且当表面基本上不含 多价阳离子污染 聚合物介质使用匹配离子多核苷酸色谱法提供多核苷酸的有效分离。 维持和储存聚合物介质的方法包括用多价阳离子结合剂进行处理。
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公开(公告)号：US06309549B1
公开(公告)日：2001-10-30
申请号：US09493779
申请日：2000-01-28
IPC分类号： B01D1508
CPC分类号： B01D15/366 , C12N15/101
摘要： Non-polar polymeric separation media, such as beads or monoliths, are suitable for chromatographic separation of mixtures of polynucleotides when the surfaces of the media are unsubstituted or substituted with a hydrocarbon group having from one to 1,000,000 carbons and when the surfaces are substantially free from mutivalent cation contamination. The polymeric media provide efficient separation of polynucleotides using Matched Ion Polynucleotide Chromatography. Methods for maintaining and storing the polymeric media include treatment with multivalent cation binding agents.
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公开(公告)号：US6066258A
公开(公告)日：2000-05-23
申请号：US183123
申请日：1998-10-30
IPC分类号： B01D15/08
CPC分类号： B01D15/366 , C12N15/101
摘要： Non-polar polymeric separation media, such as beads or monoliths, are suitable for chromatographic separation of mixtures of polynucleotides when the surfaces of the media are unsubstituted or substituted with a hydrocarbon group having from one to 1,000,000 carbons and when the surfaces are substantially free from mutivalent cation contamination. The polymeric media provide efficient separation of polynucleotides using Matched Ion Polynucleotide Chromatography. Methods for maintaining and storing the polymeric media include treatment with multivalent cation binding agents.
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公开(公告)号：US06485648B1
公开(公告)日：2002-11-26
申请号：US09696610
申请日：2000-10-25
申请人： Douglas T. Gjerde , Robert M. Haefele , Laura Bunnel , Paul D. Taylor , Kimberly A. Lamb , Satyajit Kar
发明人： Douglas T. Gjerde , Robert M. Haefele , Laura Bunnel , Paul D. Taylor , Kimberly A. Lamb , Satyajit Kar
IPC分类号： B01D1508
CPC分类号： B01D15/20 , B01D15/366 , B01J20/287 , B01J20/3425 , B01J20/3475 , G01N30/50 , G01N2030/201 , G01N2030/8827
摘要： An apparatus for effecting base pair length separations of DNA fragments by matched ion paired chromatography comprising a separation column containing separation media having non-polar DNA separation surfaces, separation solution supply means, and a separation solution conduit communicating with the separation column and the separation solution supply means, and a cleaning solution valve means positioned in the separation solution conduit for injecting cleaning solution into the separation solution conduit. A process for cleaning the non-polar DNA separation surfaces in the apparatus comprising interrupting the flow of separation solvent with a block of cleaning solution injected into the flow of separation solution passing to the column, the cleaning solution containing agent which removes accumulated residues from the non-polar surface. The cleaning solution can have an alkaline pH and contain a chelating agent such as EDTA.
摘要翻译： 一种用于通过配对离子配对色谱法实现DNA片段的碱基对长度分离的装置，其包括含有具有非极性DNA分离表面的分离介质的分离柱，分离溶液供应装置和与分离柱和分离溶液连通的分离溶液管道 供应装置和位于分离溶液管道中的清洁溶液阀装置，用于将清洁溶液注入到分离溶液导管中。 一种用于清洁设备中的非极性DNA分离表面的方法，包括用注入到通过塔的分离溶液流中的清洁溶液块中断分离溶剂的流动，该清洗溶液包含试剂从 非极性表面。 清洁溶液可以具有碱性pH并含有螯合剂如EDTA。
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公开(公告)号：US06482317B2
公开(公告)日：2002-11-19
申请号：US09828427
申请日：2001-04-05
IPC分类号： B01D1508
CPC分类号： B01D15/366 , B01D15/325 , C12N15/101
摘要： Non-polar polymeric separation media, such as beads or monoliths, are suitable for chromatographic separation of mixtures of polynucleotides when the surfaces of the media are unsubstituted or substituted with a hydrocarbon group having from one to 1,000,000 carbons and when the surfaces are substantially free from mutivalent cation contamination. The polymeric media provide efficient separation of polynucleotides using Matched Ion Polynucleotide Chromatography. Methods for maintaining and storing the polymeric media include treatment with multivalent cation binding agents.
摘要翻译： 非极性聚合物分离介质，例如珠粒或整料适用于当介质的表面未被取代或被具有1至1,000,000个碳原子的烃基取代时的多核苷酸混合物的色谱分离，并且当表面基本上不含 多价阳离子污染 聚合物介质使用匹配离子多核苷酸色谱法提供多核苷酸的有效分离。 维持和储存聚合物介质的方法包括用多价阳离子结合剂进行处理。
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公开(公告)号：US06455692B1
公开(公告)日：2002-09-24
申请号：US09698938
申请日：2000-10-26
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。
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18.发明授权Modifying double stranded DNA to enhance separations by matched ion polynucleotide chromatography 失效通过匹配的离子多核苷酸色谱法修饰双链DNA以增强分离公开(公告)号：US06210885B1
公开(公告)日：2001-04-03
申请号：US09169440
申请日：1998-10-09
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。
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19.发明授权公开(公告)号：US07225079B2
公开(公告)日：2007-05-29
申请号：US10308576
申请日：2002-12-02
CPC分类号： C12Q1/6827 , B01D15/166 , B01D15/366 , C12N15/101 , C12Q1/6869 , G01N30/30 , G01N30/96 , G01N2030/8827 , C12Q2565/137 , C12Q2527/107
摘要： In an extensive Matched Ion Polynucleotide Chromatography (MIPC) system and method, and the computer programs or software associated therewith, the system provides automated options for sample selection, mobile phase gradient selection and control, column and mobile phase temperature control, and fragment collection for a wide variety of MIPC separation processes. MIPC separation processes can be applied to effect size-based separation of DNA fragments, mutation detection, DNA fragment purification, PCR process monitoring and other novel processes. This invention is directed to the system and software which automates many of these procedures, facilitating use of the system to achieve complex separation methods. In one embodiment of the invention, a user specifies a size range of double stranded DNA fragment(s) in a mixture, the software calculates a solvent gradient to elute the fragment(s), and the system performs the chromatographic separation using the calculated gradient. In an embodiment useful in DNA mutation detection, a user specifies the base sequence of a wild type DNA molecule, the software calculates a temperature for partially denaturing heteroduplex and homoduplex molecules of the DNA in a mixture, the software calculates a solvent gradient to elute the fragments, and the system performs the chromatographic separation using the calculated gradient and temperature.
摘要翻译： 在广泛的匹配离子多核苷酸色谱（MIPC）系统和方法以及与其相关的计算机程序或软件中，系统提供用于样品选择，流动相梯度选择和控制，色谱柱和流动相温度控制以及片段收集的自动选择 各种MIPC分离过程。 可以应用MIPC分离过程来实现基于大小的DNA片段分离，突变检测，DNA片段纯化，PCR过程监控等新方法。 本发明涉及自动化许多这些程序的系统和软件，便于使用该系统来实现复杂的分离方法。 在本发明的一个实施方案中，用户指定混合物中双链DNA片段的大小范围，软件计算溶剂梯度以洗脱片段，并且系统使用计算的梯度进行色谱分离 。 在DNA突变检测中有用的实施方案中，用户指定野生型DNA分子的碱基序列，软件计算混合物中DNA的部分变性异源双链体和同源双链体分子的温度，软件计算溶剂梯度洗脱 片段，系统使用计算的梯度和温度进行色谱分离。
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公开(公告)号：US06355417B2
公开(公告)日：2002-03-12
申请号：US09039061
申请日：1998-03-13
申请人： Douglas T. Gjerde , Paul D. Taylor
发明人： Douglas T. Gjerde , Paul D. Taylor
IPC分类号： C12Q168
CPC分类号： G01N27/44721 , G01N30/8651
摘要： A method and apparatus for representing double stranded nucleic acid fragments which have been separated by a chromatographic process as an array of bands which can be accurately quantified, optimized and stored. Using, for example, a Matched Ion Polynucleotide Chromatography (MIPC) process, an analog output from a UV detector is digitized and input to a computer. The digitized signal is converted to a linear array of bands which may be displayed on a video display terminal. The intensity and/or color of a band may correlate to the amount of double stranded nucleic acid in the respective fraction or the respective double stranded nucleic acid fragment above a user selected threshold level at a corresponding point in the digitized signal. The calculated base pair length, concentration, and retention time of each band in the array of bands may be displayed in alphanumeric form.
摘要翻译： 用于表示已经通过色谱法分离的双链核酸片段的方法和装置可以被精确量化，优化和储存的条带阵列。 使用例如匹配离子多核苷酸色谱（MIPC）方法，将来自UV检测器的模拟输出数字化并输入到计算机。 数字化信号被转换成可以显示在视频显示终端上的线性阵列阵列。 条带的强度和/或颜色可以与在数字化信号的对应点处的用户选择的阈值水平之上的相应级分或相应双链核酸片段中的双链核酸的量相关。 频带阵列中每个频带的计算的碱基对长度，浓度和保留时间可以以字母数字形式显示。
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