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公开(公告)号：EP1330644A2
公开(公告)日：2003-07-30
申请号：EP01982678.3
申请日：2001-10-29
发明人： BEN-ASOULI, Yitzhak , OSMAN,Farhat
IPC分类号： G01N27/447
CPC分类号： G01N27/44743 , C07K1/26 , G01N27/44747
摘要： The present invention is directed to an apparatus for electrophoresis having a first gel matrix, adapted for performing an electrophoretic process therein, in communication with a second gel matrix, both being accommodated within a suitable housing. The housing has a first opening adapted to permit ionic communication between the first gel matrix and an external ionic buffer solution, and a second opening adapted to permit ionic communication between the second gel matrix and an external ionic buffer solution. The second gel has at least one suitable absorption material capable of retaining therein at least one target substance capable of migrating thereto from the first gel matrix when an electrophoretic process is performed in the first matrix.
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公开(公告)号：EP2404167A1
公开(公告)日：2012-01-11
申请号：EP10712797.9
申请日：2010-02-25
发明人： BEN-ASOULI, Yitzhak , OSMAN, Farhat
IPC分类号： G01N27/447
CPC分类号： G01N27/44747 , C08F220/56
摘要： Disclosed is a precast polyacrylamide gel for use in gel electrophoresis, comprising polyacrylamide and an aqueous Tris 0.04 M to 0.15 M solution, at least one first ampholyte exhibiting an isoelectric point (pI) of from 5.4 to 6.4 at a total concentration of from 0.01 M to 0.4 M; and at least one second ampholyte exhibiting an isoelectric point (pI) of from 2.5 to 3.5 to adjust the pH to between 5.5 and 7.5 at the temperature of 25°C. The precast gels can be used for electrophoresis of oligopeptides, polypeptides, oligonucleotides, and polynucleotides under denaturating or nondenaturing conditions, and exhibit long shelf-life.
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公开(公告)号：EP1330643A2
公开(公告)日：2003-07-30
申请号：EP01980886.4
申请日：2001-10-29
发明人： BEN-ASOULI, Yitzhak , OSMAN,Farhat
IPC分类号： G01N27/447
CPC分类号： G01N27/44743 , C07K1/26 , G01N27/44747
摘要： The present invention is directed to a solidified hybrid gel for use in an electrophoresis process, having a solidified first gel portion juxtaposed with a solidified second gel portion. The first gel portion is capable of accommodating therein one or more samples for electrophoresis after said first gel portion is in solidified form, and the second gel portion is adapted for enabling an electrophoresis process to be applied to such a sample that may be accommodated in said first gel portion. Thus, the hybrid gel may be provided in a precast form to users, ready for use. The invention is also directed to methods for providing such a gel, apparatuses for accommodating such a gel, and methods for carrying out electrophoresis on a sample comprised in such a gel.
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公开(公告)号：EP2183583B1
公开(公告)日：2011-06-01
申请号：EP08789846.6
申请日：2008-08-31
发明人： OSMAN, Farhat , BEN-ASOULI, Yitzhak
IPC分类号： G01N27/447 , C08F20/56 , C08F120/56 , C08F220/56 , C08L33/26
CPC分类号： G01N27/44747 , C08L33/26
摘要： Provided is a precast polyacrylamide gel for use in gel electrophoresis, having a shelf-life of at least 12 months. The gel comprises a sulfonated tertiary or secondary amine having a pKa of 6.6-8.0, ampholytes exhibiting pI between 5.4 to 6.4, Tris, and hydrochloric acid to adjust the pH to between 5.5 and 7.5.
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公开(公告)号：EP2183583A1
公开(公告)日：2010-05-12
申请号：EP08789846.6
申请日：2008-08-31
发明人： OSMAN, Farhat , BEN-ASOULI, Yitzhak
IPC分类号： G01N27/447 , C08F20/56 , C08F120/56 , C08F220/56 , C08L33/26
CPC分类号： G01N27/44747 , C08L33/26
摘要： Provided is a precast polyacrylamide gel for use in gel electrophoresis, having a shelf-life of at least 12 months. The gel comprises a sulfonated tertiary or secondary amine having a pKa of 6.6-8.0, ampholytes exhibiting pI between 5.4 to 6.4, Tris, and hydrochloric acid to adjust the pH to between 5.5 and 7.5.
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公开(公告)号：EP1285257B1
公开(公告)日：2005-07-27
申请号：EP01936753.1
申请日：2001-05-22
发明人： BEN-ASOULI, Yitzhak , OSMAN, Farhat
IPC分类号： G01N27/00
CPC分类号： G01N1/405 , B01L3/50825 , G01N2001/4016
摘要： A device having a processing chamber, particularly for electroelution and/or dialysis of a substance carried in a sample with respect to an external liquid medium. The chamber is closed at one end and has an opening at the other longitudinal end sufficiently large to permit a sample, and in particular a gel contained sample, to be inserted thereinto. The chamber has a pair of portals laterally disposed with respect to the opening. The portals are covered with a typically tubular membrane, which is sealingly fixed onto an outside surface of the housing defining the chamber, typically by means of a tubular sleeve having portals that align with the portals of the chamber, via an annular sealing arrangement. The device provides high yield recovery, saves time, and allows for relatively easy handling specially regarding loading and unloading of small volume of samples to be dialyzed or inserting the slice of gel containing the macromolecule sample. The device may be used in an open manner by partially immersing the same in the liquid medium with the opening above the liquid surface. Optionally, the device may be closed and hermetically sealed if so desired by means of a cap, and thus immersed in a liquid medium.
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公开(公告)号：EP1330643B1
公开(公告)日：2007-09-05
申请号：EP01980886.4
申请日：2001-10-29
发明人： BEN-ASOULI, Yitzhak , OSMAN,Farhat
IPC分类号： G01N27/447
CPC分类号： G01N27/44743 , C07K1/26 , G01N27/44747
摘要： The present invention is directed to a solidified hybrid gel for use in an electrophoresis process, having a solidified first gel portion juxtaposed with a solidified second gel portion. The first gel portion is capable of accommodating therein one or more samples for electrophoresis after said first gel portion is in solidified form, and the second gel portion is adapted for enabling an electrophoresis process to be applied to such a sample that may be accommodated in said first gel portion. Thus, the hybrid gel may be provided in a precast form to users, ready for use. The invention is also directed to methods for providing such a gel, apparatuses for accommodating such a gel, and methods for carrying out electrophoresis on a sample comprised in such a gel.
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公开(公告)号：EP1362101B1
公开(公告)日：2004-12-08
申请号：EP02711193.9
申请日：2002-02-11
发明人： BEN-ASOULI, Yitzhak , OSMAN, Farhat
CPC分类号： C12N15/66
摘要： The present invention relates to a method for orientation-directed construction of a construct comprising at least two nucleic acid segments of interest. In a first step, products having phosphorylated blunt ends which have been derived from the nucleic acid segments of interest are provided. In a next step, separate ligation reactions are performed to combine two segments of interest. In each reaction, the phosphorylated blunt-ended products of two different segments obtained in the first step are ligated. This ligation creates a combined ligated sequence, comprising two segments of interest attached together. The next step is enrichment of the desired orientation in the resultant product population by performing a PCR reaction using the ligated sequence obtained in the second step, as a template and specific enrichment primers directing the PCR amplification towards the desired orientation. Each combined ligated sequence is PCR amplified in a separate reaction, to create combined product having the desired orientation as well as phosphorylated blunt ends. In a fourth step, the combined product obtained in the third step is isolated and purified. If the desired construct is composed of more than two segments, the second to fourth steps may be cyclically repeated for any desired number of times, to combine all the segments of interest. The final step is performing self-ligation of the linear final isolated purified combined product obtained in the former step, creating a circular double-stranded DNA construct containing all the desired segments properly aligned and operably linked in a desired orientation.
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公开(公告)号：EP1362101A2
公开(公告)日：2003-11-19
申请号：EP02711193.9
申请日：2002-02-11
发明人： BEN-ASOULI, Yitzhak , OSMAN, Farhat
CPC分类号： C12N15/66
摘要： The present invention relates to a method for orientation-directed construction of a construct comprising at least two nucleic acid segments of interest. In a first step, products having phosphorylated blunt ends which have been derived from the nucleic acid segments of interest are provided. In a next step, separate ligation reactions are performed to combine two segments of interest. In each reaction, the phosphorylated blunt-ended products of two different segments obtained in the first step are ligated. This ligation creates a combined ligated sequence, comprising two segments of interest attached together. The next step is enrichment of the desired orientation in the resultant product population by performing a PCR reaction using the ligated sequence obtained in the second step, as a template and specific enrichment primers directing the PCR amplification towards the desired orientation. Each combined ligated sequence is PCR amplified in a separate reaction, to create combined product having the desired orientation as well as phosphorylated blunt ends. In a fourth step, the combined product obtained in the third step is isolated and purified. If the desired construct is composed of more than two segments, the second to fourth steps may be cyclically repeated for any desired number of times, to combine all the segments of interest. The final step is performing self-ligation of the linear final isolated purified combined product obtained in the former step, creating a circular double-stranded DNA construct containing all the desired segments properly aligned and operably linked in a desired orientation.
摘要翻译： 本发明涉及用于定向构建包含至少两个目的核酸片段的构建体的方法。 在第一步中，提供了具有磷酸化平端的产物，所述平端衍生自感兴趣的核酸片段。 在下一步中，进行单独的连接反应以组合两段感兴趣的部分。 在每个反应中，连接第一步中获得的两个不同区段的磷酸化平端产物。 该连接产生组合的连接序列，包括两个连在一起的感兴趣片段。 下一步是通过使用第二步中获得的连接序列进行PCR反应作为模板和引导PCR扩增朝向期望的取向的特异性富集引物，在所得产物群中富集期望的取向。 将每个组合的连接序列在单独的反应中进行PCR扩增，以产生具有所需取向以及磷酸化平端的组合产物。 在第四步中，分离并纯化第三步中得到的组合产物。 如果期望的构建体由多于两个片段组成，则第二至第四步骤可以循环重复任何期望的次数，以组合所有感兴趣的片段。 最后一步是对前一步骤中获得的线性最终分离的纯化的组合产物进行自连接，产生含有所有期望的片段的圆形双链DNA构建体，所述片段正确地对齐并且以期望的方向可操作地连接。
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公开(公告)号：EP1285257A2
公开(公告)日：2003-02-26
申请号：EP01936753.1
申请日：2001-05-22
发明人： BEN-ASOULI, Yitzhak , OSMAN, Farhat
IPC分类号： G01N27/00
CPC分类号： G01N1/405 , B01L3/50825 , G01N2001/4016
摘要： A device having a processing chamber, particularly for electroelution and/or dialysis of a substance carried in a sample with respect to an external liquid medium. The chamber is closed at one end and has an opening at the other longitudinal end sufficiently large to permit a sample, and in particular a gel contained sample, to be inserted thereinto. The chamber has a pair of portals laterally disposed with respect to the opening. The portals are covered with a typically tubular membrane, which is sealingly fixed onto an outside surface of the housing defining the chamber, typically by means of a tubular sleeve having portals that align with the portals of the chamber, via an annular sealing arrangement. The device provides high yield recovery, saves time, and allows for relatively easy handling specially regarding loading and unloading of small volume of samples to be dialyzed or inserting the slice of gel containing the macromolecule sample. The device may be used in an open manner by partially immersing the same in the liquid medium with the opening above the liquid surface. Optionally, the device may be closed and hermetically sealed if so desired by means of a cap, and thus immersed in a liquid medium.
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