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1.发明授权公开(公告)号：US06399305B1
公开(公告)日：2002-06-04
申请号：US09588950
申请日：2000-06-07
申请人： Yoshihiko Makino , Yoshihiko Abe , Makoto Takagi , Shigeori Takenaka , Kenichi Yamashita , Masashi Ogawa
发明人： Yoshihiko Makino , Yoshihiko Abe , Makoto Takagi , Shigeori Takenaka , Kenichi Yamashita , Masashi Ogawa
IPC分类号： C12Q168
CPC分类号： C12Q1/6825 , Y10T436/143333 , C12Q2563/173 , C12Q2563/113
摘要： A method of analyzing a nucleic acid fragment sample to judge whether the nucleic acid fragment sample is uncomplementary, partly complementary or complementary to a DNA fragment in its specific base sequence is conducted by the steps of: bringing an aqueous solution of the nucleic acid fragment sample into contact with a DNA chip having an electroconductive substrate and the DNA fragment fixed onto the substrate in the presence of an electrochemical thread intercalator; measuring an electric current flowing from or to the electroconductive substrate along the DNA fragment under application of a potential to the substrate; and comparing the electric current measured above with a referential electric current which is prepared employing a DNA chip equivalent to the above DNA chip, the intercalator, and an aqueous solution of a nucleic acid fragment which is complementary to the DNA fragment of the DNA chip.
摘要翻译： 分析核酸片段样品以判断核酸片段样品是否与其特异性碱基序列中的DNA片段不互补，部分互补或互补的方法通过以下步骤进行：使核酸片段样品的水溶液 与具有导电性基体的DNA芯片接触，并且在电化学线性嵌入剂的存在下固定在基材上的DNA片段;测量在施加电位下沿着DNA片段向基片流向或流到导电基底的电流 ; 并使用与上述DNA芯片相当的DNA芯片，嵌入剂和与DNA芯片的DNA片段互补的核酸片段的水溶液制备的参考电流来比较上述电流。
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公开(公告)号：US06864055B2
公开(公告)日：2005-03-08
申请号：US09887625
申请日：2001-06-22
申请人： Yoshihiko Makino , Yoshihiko Abe , Masashi Ogawa , Makoto Takagi , Shigeori Takenaka , Kenichi Yamashita
发明人： Yoshihiko Makino , Yoshihiko Abe , Masashi Ogawa , Makoto Takagi , Shigeori Takenaka , Kenichi Yamashita
IPC分类号： G01N33/53 , C07B61/00 , C07K14/00 , C12M1/00 , C12N15/09 , C12Q1/68 , G01N27/416 , G01N33/566 , B01D57/02 , C12P19/34 , G01N30/02
CPC分类号： C12Q1/6825 , C07K14/003 , C12Q1/6837 , C40B40/00 , C12Q2565/501 , C12Q2565/102 , C12Q2565/607
摘要： A method of detecting nucleic acid fragments in plural samples is performed by the steps of: attaching an electroconductive label to nucleic acid fragments in one sample and attaching a different electroconductive label to nucleic acid fragments in another sample; preparing a mixture of these samples; spotting the mixture on an electroconductive microarray having plural electrodes onto which probe molecules complementary to the nucleic acid fragments are fixed, so that hybridization between the nucleic acid fragments and the probe molecules on the electroconductive microarray can proceed to form hybrid structures; applying to the electrode an electric potential corresponding to the oxidation-reduction potential of the former label and detecting on the electrode an electric current; applying to the electrode an electric potential corresponding to the oxidation-reduction potential of the latter label and detecting on the electrode an electric current; and comparing the electric current detected in the former detecting procedure and that detected in the latter detecting procedure.
摘要翻译： 通过以下步骤进行检测多个样品中的核酸片段的方法：将导电标记物附着在一个样品中的核酸片段上，并将不同的导电标记连接到另一个样品中的核酸片段; 准备这些样品的混合物; 在具有多个电极的导电微阵列上将该混合物点样，在该导电微阵列上固定有与核酸片段互补的探针分子，使得导电微阵列上的核酸片段和探针分子之间的杂交可以进行以形成杂化结构; 向所述电极施加与所述前标签的氧化还原电位相对应的电位，并在所述电极上检测电流; 向所述电极施加与后一标签的氧化还原电位相对应的电位，并在所述电极上检测电流; 并比较前一检测程序检测的电流和后一检测程序中检测的电流。
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公开(公告)号：US06713255B1
公开(公告)日：2004-03-30
申请号：US09589221
申请日：2000-06-07
申请人： Yoshihiko Makino , Yoshihiko Abe , Makoto Takagi , Shigeori Takenaka , Kenichi Yamashita , Masashi Ogawa
发明人： Yoshihiko Makino , Yoshihiko Abe , Makoto Takagi , Shigeori Takenaka , Kenichi Yamashita , Masashi Ogawa
IPC分类号： C12Q168
CPC分类号： C07K14/003 , B01J19/0046 , B01J2219/00529 , B01J2219/00585 , B01J2219/00596 , B01J2219/00605 , B01J2219/00608 , B01J2219/0061 , B01J2219/00612 , B01J2219/00617 , B01J2219/00626 , B01J2219/00637 , B01J2219/00641 , B01J2219/00659 , B01J2219/00722 , B01J2219/00729 , C07B2200/11 , C07H21/00 , C12Q1/6837 , C40B40/06 , C12Q2565/607
摘要： A DNA chip (or PNA chip) composed of a solid carrier and plural DNA fragments (or PNA fragments) fixed onto is the solid carrier at each one end, wherein a plurality of short chain spacer molecules having a hydrophilic moiety at each one end are fixed at each another end onto a surface of the solid carrier having no DNA fragments (or no PNA fragments) on its surface is effective for high sensitive quantitative analysis of a nucleic acid fragment complementary to the DNA fragment (or PNA fragment).
摘要翻译： 由固体载体和固定在其上的多个DNA片段（或PNA片段）组成的DNA芯片（或PNA芯片）是每一端的固体载体，其中在每一端具有亲水部分的多个短链间隔基分子是 固定在其表面上没有DNA片段（或没有PNA片段）的固体载体的表面上对于与DNA片段（或PNA片段）互补的核酸片段的高敏感性定量分析是有效的。
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公开(公告)号：US06506567B2
公开(公告)日：2003-01-14
申请号：US09774895
申请日：2001-01-31
IPC分类号： C12Q168
CPC分类号： C07D471/06 , C12Q1/6816 , Y10T436/143333 , C12Q2563/173
摘要： A water-soluble fluorescent intercalator compound having the formula: F—La—X F is a fluorescent moiety, X is a divalent cyclic group, and La is a linking group, and at least one of X and La has a site imparting water solubility to the compound or a site that is convertible into a site imparting water solubility to the compound is favorably employable as a fluorescent intercalator in a method for fluorometrically detecting complementary DNA fragments.
摘要翻译： 具有式：F的水溶性荧光嵌入剂化合物是荧光部分，X是二价环状基团，La是连接基团，X和La中的至少一个具有赋予化合物水溶性的部位或 可以转化为赋予化合物水溶性的部位的部位在荧光测定互补DNA片段的方法中有利地用作荧光嵌入剂。
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公开(公告)号：US06368807B2
公开(公告)日：2002-04-09
申请号：US09733365
申请日：2000-12-08
IPC分类号： C12Q168
CPC分类号： C07F17/02
摘要： A compound of the following formula: Ea—La—X—Lb—Eb in which each of Ea and Eb independently is a group having oxidation-reduction activity and having a conjugated system in its group; X is a divalent cyclic group; and each of La and Lb independently is a group which does not form a conjugated system in combination with the conjugated system of each of Ea and Eb and at least one of which has a site imparting water solubility to the compound or a site that is convertible into a site imparting water solubility to the compound, is favorably employable as an electroconductive threading intercalator in an electrochemical method for detecting complementary DNA fragments.
摘要翻译： 下式的化合物：其中Ea和Eb各自独立地是具有氧化还原活性的基团并且在其组中具有共轭体系; X是二价环状基团; 独立地，La和Lb各自是不与Ea和Eb各自的共轭体系结合形成共轭体系的基团，并且其中至少一个具有赋予化合物或可转化位点水溶性的位点 成为赋予化合物水溶性的部位有利地用作检测互补DNA片段的电化学方法中的导电穿线嵌入剂。
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公开(公告)号：US07132266B2
公开(公告)日：2006-11-07
申请号：US10292725
申请日：2002-11-12
申请人： Yoshihiko Makino , Yoshihiko Abe
发明人： Yoshihiko Makino , Yoshihiko Abe
CPC分类号： C12Q1/6825
摘要： A method of analyzing a target nucleic acid fragment is provided that is capable of being performed simply and promptly, by anyone, using a compact device without requirement of specific technique or complex operation. The method comprises timely detection of a double-stranded nucleic acid fragment formed during the polymerase extension reaction due to a specific base sequence of the target nucleic acid fragment as increase of an electrochemical response under the existence of an electrochemically active intercalator; and the kit utilizes the method.
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公开(公告)号：US07321831B2
公开(公告)日：2008-01-22
申请号：US10231242
申请日：2002-08-30
申请人： Yoshihiko Makino , Yoshihiko Abe
发明人： Yoshihiko Makino , Yoshihiko Abe
摘要： It is an object of the present invention to provide a nucleic acid fragment-fixed electrode wherein a probe nucleic acid fragment is fixed on the electrode stably and in an amount-controlled manner. The present invention provides a nucleic acid fragment-fixed electrode wherein a nucleic acid fragment is fixed on the surface of a multi-component self-assembled monolayer of two or more different components which is formed on the electrode, by a covalent bond via a bifunctional linking molecule.
摘要翻译： 本发明的目的是提供一种核酸片段固定电极，其中探针核酸片段稳定地并以量控制的方式固定在电极上。 本发明提供了一种核酸片段固定电极，其中核酸片段固定在多组分自组装单层的两个或多个不同组分的表面上，该组分形成在电极上，通过共价键通过双功能 连接分子。
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8.发明申请公开(公告)号：US20060068491A1
公开(公告)日：2006-03-30
申请号：US11227245
申请日：2005-09-16
申请人： Yoshihiko Makino , Yoshiki Sakaino , Yukio Sudo , Yoshihiko Abe
发明人： Yoshihiko Makino , Yoshiki Sakaino , Yukio Sudo , Yoshihiko Abe
CPC分类号： B01L3/502753 , B01L3/502707 , B01L3/502715 , B01L2200/0631 , B01L2300/0816 , B01L2300/0867 , B01L2400/0406 , B01L2400/0418 , B01L2400/0487
摘要： A microdevice for performing a method for separating and purifying a nucleic acid, the microdevice comprising: at least one opening; and at least one channel for passing a sample solution, wherein the method comprises: (A) a step of bringing a nucleic acid-containing sample solution into contact with a nucleic acid-adsorbing support having a function of adsorbing a nucleic acid; (B) a step of washing the nucleic acid-adsorbing support with a washing solution in a state of a nucleic acid being adsorbed to the support; and (C) a step of desorbing the nucleic acid from the nucleic acid-adsorbing support by a recovering solution, thereby purifying the nucleic acid; an apparatus for utilizing the microdevice; and a reagent kit for use in the microdevice.
摘要翻译： 一种用于进行分离和纯化核酸的方法的微型装置，所述微型装置包括：至少一个开口; 和用于通过样品溶液的至少一个通道，其中所述方法包括：（A）使含核酸样品溶液与具有吸附核酸功能的核酸吸附载体接触的步骤; （B）在被吸附在所述载体上的核酸的状态下用洗涤液洗涤所述核酸吸附载体的工序; 和（C）通过回收溶液从核酸吸附载体解吸核酸的步骤，从而纯化核酸; 一种利用微型装置的装置; 和用于微型装置的试剂盒。
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9.发明授权
公开(公告)号：US5130258A
公开(公告)日：1992-07-14
申请号：US643831
申请日：1991-01-18
申请人： Yoshihiko Makino , Masashi Ogawa
发明人： Yoshihiko Makino , Masashi Ogawa
IPC分类号： G01N21/27 , G01N21/78 , G01N33/487 , G01N33/49 , G01N33/52
CPC分类号： G01N33/525 , G01N21/272 , G01N33/491 , G01N21/78 , Y10S435/805 , Y10S435/81 , Y10S436/805 , Y10S436/808 , Y10S436/824 , Y10T436/255
摘要： A method of quantitatively analyzing an analyte contained in a whole blood sample, wherein a dry multi-layered analysis element is used. The method provides particular merits when the used multi-layered analysis element has no light-shielding layer which is interposed, in the conventional analysis element, between a coloring reagent layer and a blood cell separating layer, so that red coloring matters of blood cells are detected from the support side during the step of measuring the optical density of the reflected light. After the optical density due to coloring matters of blood cells trapped by the blood cell separating layer has reached a constant level or background density, the changing rate in optical density is measured and then the measured changing rate is converted to the corresponding content or activity of the analyte through a colorimetric operation, or the total change in optical density is measured and then the substantially constant background density is subtracted therefrom to know the change in optical density caused by a coloring dye or like material formed in the coloring reagent layer in the presence of the analyte, followed by a similar conversion operation performed on the basis of the principle of colorimetry, to determine the content or activity of the analyte.
摘要翻译： 一种定量分析全血样品中包含的分析物的方法，其中使用干燥的多层分析元件。 当使用的多层分析元件在传统分析元件中没有遮光层在着色剂层和血细胞分离层之间时，该方法提供了特别的优点，使得血细胞的红色着色物质为 在测量反射光的光密度的步骤期间从支撑侧检测到。 在由血细胞分离层捕获的血细胞的着色物质的光密度达到一定水平或背景浓度后，测定光密度的变化率，然后将测定的变化率转换成相应的含量或活性 通过比色操作的分析物或光密度的总变化被测量，然后从其中减去基本上恒定的背景密度，以了解由存在着色剂层中形成的着色染料或类似材料引起的光密度的变化 的分析物，然后根据比色法原理进行类似的转化操作，以确定分析物的含量或活性。
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公开(公告)号：US4900416A
公开(公告)日：1990-02-13
申请号：US243975
申请日：1988-09-13
申请人： Yoshihiko Makino , Masashi Ogawa
发明人： Yoshihiko Makino , Masashi Ogawa
IPC分类号： G01N27/26 , G01N27/447
CPC分类号： G01N27/44704 , G01N27/44747
摘要： An electrophoretic method comprises the steps of positioning a sample, which contains nucleic acid fragments having different numbers of bases, in an electrophoresis medium composed of an electrophoresis gel medium containing an aqueous poly(meth)acrylamide gel prepared by cross-linking polymerization of a (meth)acrylamide compound and a cross-linking agent in the presence of water and a compound having at least one carbamoyl group as a denaturing agent (or modifier), and applying pulsed electric fields to the electrophoresis medium in two directions, thereby to move the nucleic acid fragments in the presence of molecular sieve effects and to separate the nucleic acid fragments.
摘要翻译： 电泳方法包括以下步骤：将含有不同碱基数的核酸片段的样品置于电泳介质中，所述电泳介质由电泳凝胶介质组成，所述电泳凝胶介质含有聚（甲基）丙烯酰胺凝胶水溶液， 甲基）丙烯酰胺化合物和交联剂在水和具有至少一个氨基甲酰基的化合物作为变性剂（或改性剂）的存在下进行，并且在两个方向上向电泳介质施加脉冲电场，从而使 核酸片段在分子筛效应的存在下分离核酸片段。
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