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公开(公告)号：US07687234B2
公开(公告)日：2010-03-30
申请号：US11011265
申请日：2004-12-13
申请人： Ting Chen , Xing Yang , R. Erik Holmlin
发明人： Ting Chen , Xing Yang , R. Erik Holmlin
CPC分类号： G01N33/5438 , B01J19/0046 , B01J2219/00432 , B01J2219/00441 , B01J2219/00497 , B01J2219/00527 , B01J2219/00585 , B01J2219/00596 , B01J2219/00653 , B01J2219/00677 , B01J2219/00722 , B01J2219/00725 , B01J2219/00729 , C12Q1/6825 , C12Q1/6837 , C12Q2565/501 , C12Q2565/607
摘要： Methods for conducting an assay to detect nucleic acid hybridization are disclosed. In particular, the fabrication of a carbon electrode suited to nucleic acid hybridization detection is described. In some preferred embodiments, a micro array of carbon electrodes is constructed using photolithography. The final step in the photolithography process involves developing a solder mask with an alkaline solution. The alkaline solution oxidizes the carbon surface producing surface carboxylic acid groups. The surface carboxylic acid groups are reacted with EDC or DCC and NHS to produce NHS esters. Immobilization of NEUTRAVIDIN® onto the electrode surface is effected by reaction with the surface NHS ester groups. A biotinylated probe DNA molecule is then attached to the electrode via binding between the biotin group and the immobilized NEUTRAVIDIN®. The resulting electrode can be used to detect hybridization between the probe DNA and complementary target DNA.
摘要翻译： 公开了用于进行测定以检测核酸杂交的方法。 特别地，描述了适合于核酸杂交检测的碳电极的制造。 在一些优选实施例中，使用光刻法构建微电极阵列的碳电极。 光刻工艺的最后一步包括用碱性溶液开发焊接掩模。 碱性溶液氧化碳表面产生表面羧酸基团。 表面羧酸基团与EDC或DCC和NHS反应生成NHS酯。 将NEUTRAVIDIN®固定在电极表面上是通过与表面NHS酯基团的反应进行的。 然后通过生物素基团和固定化的NEUTRAVIDIN之间的结合将生物素化的探针DNA分子连接到电极。 所得电极可用于检测探针DNA和互补靶DNA之间的杂交。
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2.发明申请ELECTROCHEMICAL METHOD TO MEASURE DNA ATTACHMENT TO AN ELECTRODE SURFACE IN THE PRESENCE OF MOLECULAR OXYGEN 失效在分子氧存在下测量DNA连接到电极表面的电化学方法公开(公告)号：US20100018876A1
公开(公告)日：2010-01-28
申请号：US12569866
申请日：2009-09-29
CPC分类号： C12Q1/6825 , C12Q1/682 , C12Q1/6827 , C12Q1/6837 , C12Q2565/607 , C12Q2563/137 , C12Q2533/107 , C12Q2531/125 , C12Q2521/301 , C12Q2525/179 , C12Q2565/501 , C12Q2531/137 , C12Q2527/107
摘要： The present disclosure provides methods and compositions for conducting an assay to detect a polynucleotide. In particular, ruthenium complexes having reduction potentials that do not coincide with the reduction potential of molecular oxygen are disclosed and amperometric techniques for their use are described. In preferred embodiments, the ruthenium complex is ruthenium (III) pentaamine pyridine and the polynucleotide that is detected is DNA. Further, techniques for enhancing detectable contrast between hybridized and unhybridized nucleic acids are disclosed. In particular, the use of elongated target strands as well as the use of uncharged probe strands are discussed.
摘要翻译： 本公开提供用于进行测定以检测多核苷酸的方法和组合物。 特别地，公开了具有与分子氧的还原电位不一致的还原电位的钌络合物，并且描述了它们的使用的电流分析技术。 在优选的实施方案中，钌络合物是钌（III）五胺吡啶，并且所检测的多核苷酸是DNA。 此外，公开了用于增强杂交和未杂交核酸之间的可检测对比度的技术。 特别地，讨论了使用细长的靶股以及使用不带电的探针股线。
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3.发明授权Electrochemical method to measure DNA attachment to an electrode surface in the presence of molecular oxygen 失效在分子氧存在下测量DNA附着到电极表面的电化学方法公开(公告)号：US07258978B2
公开(公告)日：2007-08-21
申请号：US10429291
申请日：2003-05-02
CPC分类号： C12Q1/6825 , C12Q1/682 , C12Q1/6827 , C12Q1/6837 , C12Q2565/607 , C12Q2563/137 , C12Q2533/107 , C12Q2531/125 , C12Q2521/301 , C12Q2525/179 , C12Q2565/501 , C12Q2531/137 , C12Q2527/107
摘要： The present disclosure provides methods and compositions for conducting an assay to detect nucleic acid hybridization in the presence of oxygen. In particular, ruthenium complexes having a reduction potential that does not coincide with the reduction potential of molecular oxygen are disclosed and amperometric techniques for their use are described. In preferred embodiments, the ruthenium complex is ruthenium (III) pentaamine pyridine and the nucleic acid hybridization event that is detected is DNA hybridization. Further, techniques for enhancing detectable contrast between hybridized and unhybridized nucleic acids are disclosed. In particular, the use of elongated target strands as well as the use of uncharged probe strands are discussed.
摘要翻译： 本公开提供用于进行测定以在氧存在下检测核酸杂交的方法和组合物。 特别地，公开了具有与分子氧的还原电位不一致的还原电位的钌络合物，并且描述了它们的使用的电流分析技术。 在优选的实施方案中，钌络合物是钌（III）五胺吡啶，并且所检测的核酸杂交事件是DNA杂交。 此外，公开了用于增强杂交和未杂交核酸之间的可检测对比度的技术。 特别地，讨论了使用细长的靶股以及使用不带电的探针股线。
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4.发明授权Electrochemical method to measure DNA attachment to an electrode surface in the presence of molecular oxygen 失效在分子氧存在下测量DNA附着到电极表面的电化学方法公开(公告)号：US07879554B2
公开(公告)日：2011-02-01
申请号：US12569866
申请日：2009-09-29
CPC分类号： C12Q1/6825 , C12Q1/682 , C12Q1/6827 , C12Q1/6837 , C12Q2565/607 , C12Q2563/137 , C12Q2533/107 , C12Q2531/125 , C12Q2521/301 , C12Q2525/179 , C12Q2565/501 , C12Q2531/137 , C12Q2527/107
摘要： The present disclosure provides methods and compositions for conducting an assay to detect a polynucleotide. In particular, ruthenium complexes having reduction potentials that do not coincide with the reduction potential of molecular oxygen are disclosed and amperometric techniques for their use are described. In preferred embodiments, the ruthenium complex is ruthenium (III) pentaamine pyridine and the polynucleotide that is detected is DNA. Further, techniques for enhancing detectable contrast between hybridized and unhybridized nucleic acids are disclosed. In particular, the use of elongated target strands as well as the use of uncharged probe strands are discussed.
摘要翻译： 本公开提供用于进行测定以检测多核苷酸的方法和组合物。 特别地，公开了具有与分子氧的还原电位不一致的还原电位的钌络合物，并且描述了它们的使用的电流分析技术。 在优选的实施方案中，钌络合物是钌（III）五胺吡啶，并且所检测的多核苷酸是DNA。 此外，公开了用于增强杂交和未杂交核酸之间的可检测对比度的技术。 特别地，讨论了使用细长的靶股以及使用不带电的探针股线。
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5.发明授权Electrochemical method to measure DNA attachment to an electrode surface in the presence of molecular oxygen 有权在分子氧存在下测量DNA附着到电极表面的电化学方法公开(公告)号：US07615348B2
公开(公告)日：2009-11-10
申请号：US11836046
申请日：2007-08-08
CPC分类号： C12Q1/6825 , C12Q1/682 , C12Q1/6827 , C12Q1/6837 , C12Q2565/607 , C12Q2563/137 , C12Q2533/107 , C12Q2531/125 , C12Q2521/301 , C12Q2525/179 , C12Q2565/501 , C12Q2531/137 , C12Q2527/107
摘要： The present disclosure provides methods and compositions for conducting an assay to detect a polynucleotide. In particular, ruthenium complexes having reduction potentials that do not coincide with the reduction potential of molecular oxygen are disclosed and amperometric techniques for their use are described. In preferred embodiments, the ruthenium complex is ruthenium (III) pentaamine pyridine and the polynucleotide that is detected is DNA. Further, techniques for enhancing detectable contrast between hybridized and unhybridized nucleic acids are disclosed. In particular, the use of elongated target strands as well as the use of uncharged probe strands are discussed.
摘要翻译： 本公开提供用于进行测定以检测多核苷酸的方法和组合物。 特别地，公开了具有与分子氧的还原电位不一致的还原电位的钌络合物，并且描述了它们的使用的电流分析技术。 在优选的实施方案中，钌络合物是钌（III）五胺吡啶，并且所检测的多核苷酸是DNA。 此外，公开了用于增强杂交和未杂交核酸之间的可检测对比度的技术。 特别地，讨论了使用细长的靶股以及使用不带电的探针股线。
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