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    Multiplex polynucleotide capture methods and compositions
    2.
    发明授权
    Multiplex polynucleotide capture methods and compositions 有权

    公开(公告)号：US06514699B1

    公开(公告)日：2003-02-04

    申请号：US09593312

    申请日：2000-06-13

    申请人： Roger A. O'Neill ,  Jer-Kang Chen ,  Claudia Chiesa ,  George Fry

    发明人： Roger A. O'Neill ,  Jer-Kang Chen ,  Claudia Chiesa ,  George Fry

    IPC分类号： C12Q168

    CPC分类号： C12Q1/6874 ,  C12Q2565/514 ,  C12Q2537/157 ,  C12Q2525/161 ,  C12Q2565/629

    摘要： The invention relates to methods and compositions for simultaneously generating a plurality of polynucleotide sequencing ladders or PCR amplification products. Each sequencing ladder is generated from a recoverable primer, i.e., an oligonucleotide primer comprising a recovery tag. The recovery tag may be an oligonucleotide. Each sequencing ladder has a unique recovery tag. After the generation of the multiple sequencing ladders, the different sequencing ladders are separated from one another, i.e., purified, by binding to recovery tag binding compounds that have been immobilized on one or more solid supports. The recovery tag binding compounds are immobilized on the solid support in an addressable manner, i.e., the recovery tag binding compounds have distinct locations on the solid support. The binding of the sequencing ladders to the recovery tag binding compounds serves to separate the different polynucleotide sequencing ladders present in a given solution. The separated sequencing ladders may then be released from the immobilized recovery tag binding compounds and subsequently resolved into individual components of the sequencing ladders or PCR products. The subject methods of separating sequencing ladders simultaneously generated in the same vessel may readily be adapted to separate a plurality of simultaneously generated polynucleotide amplification products. Other aspects of the invention are kits for the generation or recovery of a plurality of polynucleotide sequencing ladders or amplification products. The kits comprise a plurality of recoverable primers having unique recovery tags. Preferably, the recovery tags are polynucleotides that have substantially the same denaturation temperature when bound to appropriate recovery tag binding compounds, i.e., the primers comprise an integrated set. The kits may further comprise recovery tag binding compounds. The kits may further comprise labeled chain terminators. Other aspects of the invention include polynucleotide recovery devices.

    Electrophoretic nucleic acid purification method
    4.
    发明授权
    Electrophoretic nucleic acid purification method 失效
    电泳核酸纯化方法

    公开(公告)号：US6146511A

    公开(公告)日：2000-11-14

    申请号：US016531

    申请日：1998-01-30

    申请人： Gary Slater ,  J. William Efcavitch ,  Guy Drouin ,  Pascal Mayer ,  Jean Rousseau ,  Hong Yan Zhou ,  Claudia Chiesa ,  Robert Ruhfel ,  Roger O'Neill

    发明人： Gary Slater ,  J. William Efcavitch ,  Guy Drouin ,  Pascal Mayer ,  Jean Rousseau ,  Hong Yan Zhou ,  Claudia Chiesa ,  Robert Ruhfel ,  Roger O'Neill

    IPC分类号： B01D57/02 ,  B03C5/00 ,  C07H21/02 ,  C07H21/04 ,  C12N15/09 ,  C12N15/10 ,  G01N27/447 ,  G01N27/26

    CPC分类号： C12N15/101 ,  G01N27/447

    摘要： An electrophoretic method for purifying a nucleic acid sample is disclosed. The method generally comprises the steps of (1) providing a nucleic acid sample comprising a desired nucleic acid and one or more contaminants, (2) providing an electrophoresis matrix having a loading well and a recovery well formed therein, (3) placing the nucleic acid sample into the loading well, (4) performing a first electrophoresis comprising electrophoresing the nucleic acid sample for a first time effective to transport the desired nucleic acid out of the loading well and into the electrophoresis matrix; and (5) performing a second electrophoresis comprising electrophoresing the nucleic acid sample for a second time effective to transport the desired nucleic acid out of the electrophoresis matrix and into the recovery well. According to the method, the first and second electrophoresis steps are effective to substantially reduce the concentration of contaminants relative to the concentration of desired nucleic acid in the nucleic acid sample, thereby producing a purified nucleic acid. In the method, the loading and recovery wells may be the same or different, and the electric fields may be DC or alternating. Also disclosed is a preparative electrophoresis method employing an alternating electrical field.

    摘要翻译： 公开了一种用于纯化核酸样品的电泳方法。 该方法通常包括以下步骤：（1）提供包含所需核酸和一种或多种污染物的核酸样品，（2）提供具有载体孔的电泳基质和其中形成的回收孔，（3） （4）进行第一次电泳，其中包括使核酸样品首次有效地将所需核酸转运到载体孔中并进入电泳基质中; 包括进行第二次电泳，其包括将核酸样品电泳第二次，有效将所需核酸转运到电泳基质内并进入回收井。 根据该方法，第一和第二电泳步骤对于相对于核酸样品中所需核酸浓度显着降低污染物的浓度是有效的，从而产生纯化的核酸。 在该方法中，加载和回收井可以相同或不同，并且电场可以是直流或交替的。 还公开了采用交流电场的制备电泳方法。

    Multiplex polynucleotide capture methods and compositions
    5.
    发明授权
    Multiplex polynucleotide capture methods and compositions 失效

    公开(公告)号：US6124092A

    公开(公告)日：2000-09-26

    申请号：US873437

    申请日：1997-06-12

    申请人： Roger A. O'Neill ,  Jer-Kang Chen ,  Claudia Chiesa ,  George Fry

    发明人： Roger A. O'Neill ,  Jer-Kang Chen ,  Claudia Chiesa ,  George Fry

    IPC分类号： C12N15/09 ,  C12Q1/68 ,  C07H21/02 ,  C07H21/04 ,  C12P19/34

    CPC分类号： C12Q1/6874

    摘要： The invention relates to methods and compositions for simultaneously generating a plurality of polynucleotide sequencing ladders or PCR amplification products. Each sequencing ladder is generated from a recoverable primer, i.e., an oligonucleotide primer comprising a recovery tag. The recovery tag may be an oligonucleotide. Each sequencing ladder has a unique recovery tag. After the generation of the multiple sequencing ladders, the different sequencing ladders are separated from one another, i.e., purified, by binding to recovery tag binding compounds that have been immobilized on one or more solid supports. The recovery tag binding compounds are immobilized on the solid support in an addressable manner, i.e., the recovery tag binding compounds have distinct locations on the solid support. The binding of the sequencing ladders to the recovery tag binding compounds serves to separate the different polynucleotide sequencing ladders present in a given solution. The separated sequencing ladders may then be released from the immobilized recovery tag binding compounds and subsequently resolved into individual components of the sequencing ladders or PCR products. The subject methods of separating sequencing ladders simultaneously generated in the same vessel may readily be adapted to separate a plurality of simultaneously generated polynucleotide amplification products. Other aspects of the invention are kits for the generation or recovery of a plurality of polynucleotide sequencing ladders or amplification products. The kits comprise a plurality of recoverable primers having unique recovery tags. Preferably, the recovery tags are polynucleotides that have substantially the same denaturation temperature when bound to appropriate recovery tag binding compounds, i.e., the primers comprise an integrated set. The kits may further comprise recovery tag binding compounds. The kits may further comprise labeled chain terminators. Other aspects of the invention include polynucleotide recovery devices.

    Method and silicate composition for conditioning silica surfaces
    7.
    发明授权
    Method and silicate composition for conditioning silica surfaces 失效
    用于调节二氧化硅表面的方法和硅酸盐组合物

    公开(公告)号：US06297009B1

    公开(公告)日：2001-10-02

    申请号：US09005075

    申请日：1998-01-09

    申请人： David M. Demorest ,  Stephen E. Moring ,  Claudia Chiesa

    发明人： David M. Demorest ,  Stephen E. Moring ,  Claudia Chiesa

    IPC分类号： C12Q168

    CPC分类号： G01N27/44752 ,  G01N30/6052 ,  G01N2030/285

    摘要： Disclosed is a method for increasing the electroosmotic flow rate available for a silica surface. In the method, there is provided an electrophoretic channel which is defined by one or more silica surfaces. The surface(s) are contacted with an alkaline aqueous solution containing a solubilized silicate-monovalent metal complex in an amount effective to increase the acidity of the silica surface(s), as evidenced by a reduction in the average bulk pKa of the surface(s). The achieved increase in acidity is greater than would be obtained using an otherwise identical solution lacking said silicate. In one preferred embodiment, the monovalent metal used in the solution is Li+, Na+, or K+. Also disclosed is a method for increasing the acidity of a silica surface, by contacting the surface with an alkaline aqueous solution of the type noted above.

    摘要翻译： 公开了一种增加可用于二氧化硅表面的电渗流速的方法。 在该方法中，提供了由一个或多个二氧化硅表面限定的电泳通道。 表面与含有增溶的硅酸盐 - 一价金属络合物的碱性水溶液以有效提高二氧化硅表面酸度的量接触，这通过表面平均体积pKa的降低来证明（ s）。 实现的酸度增加大于使用其它相同的不含所述硅酸盐的溶液所获得的增加。 在一个优选的实施方案中，溶液中使用的一价金属是Li +，Na +或K +。 还公开了通过使表面与上述类型的碱性水溶液接触来提高二氧化硅表面的酸度的方法。

    Method and silicate composition for conditioning silica surfaces
    8.
    发明授权
    Method and silicate composition for conditioning silica surfaces 失效
    用于调节二氧化硅表面的方法和硅酸盐组合物

    公开(公告)号：US5578179A

    公开(公告)日：1996-11-26

    申请号：US501674

    申请日：1995-07-12

    申请人： David M. Demorest ,  Stephen E. Moring ,  Claudia Chiesa

    发明人： David M. Demorest ,  Stephen E. Moring ,  Claudia Chiesa

    IPC分类号： G01N27/447 ,  G01N30/28 ,  G01N30/60 ,  G01N27/26

    CPC分类号： G01N27/44752 ,  G01N2030/285 ,  G01N30/6052

    摘要： Disclosed is a method for increasing the electro-osmotic flow rate available for a silica surface. In the method, there is provided an electrophoretic channel which is defined by one or more silica surfaces. The surface(s) are contacted with an alkaline aqueous solution containing a solubilized silicate-monovalent metal complex in an amount effective to increase the acidity of the silica surface(s), as evidenced by a reduction in the average bulk pKa of the surface(s). The achieved increase in acidity is greater than would be obtained using an otherwise identical solution lacking said silicate. In one preferred embodiment, the monovalent metal used in the solution is Li.sup.+, Na.sup.+, or K.sup.+. Also disclosed is a method for increasing the acidity of a silica surface, by contacting the surface with an alkaline aqueous solution of the type noted above.

    摘要翻译： 公开了一种增加可用于二氧化硅表面的电渗流速的方法。 在该方法中，提供了由一个或多个二氧化硅表面限定的电泳通道。 表面与含有增溶的硅酸盐 - 一价金属络合物的碱性水溶液以有效提高二氧化硅表面酸度的量接触，这通过表面平均体积pKa的降低来证明（ s）。 实现的酸度增加大于使用其它相同的不含所述硅酸盐的溶液所获得的增加。 在一个优选的实施方案中，溶液中使用的一价金属是Li +，Na +或K +。 还公开了通过使表面与上述类型的碱性水溶液接触来提高二氧化硅表面的酸度的方法。