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公开(公告)号：US07049104B2
公开(公告)日：2006-05-23
申请号：US10353033
申请日：2003-01-29
申请人： Hideki Kambara , Zheng Ping Li , Kazunori Okano , Keiichi Nagai
发明人： Hideki Kambara , Zheng Ping Li , Kazunori Okano , Keiichi Nagai
CPC分类号： C12Q1/6876 , C12Q1/6858 , C12Q2600/156 , C12Q2535/125 , C12Q2525/186
摘要： The present invention relates to a quick and simple method for genetic testing, particularly for SNP testing, using two kinds of primers which are complementary to a mutant target and a wild-type target, respectively, and different in length or migration speed in electrophoresis. These probes are allowed to hybridize with targets, fluorophore-tagged nucleotides are attached to the primers, and electrophoresis is carried out for discriminatory detection.
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公开(公告)号：US5356776A
公开(公告)日：1994-10-18
申请号：US942470
申请日：1992-09-09
CPC分类号： G01N27/44726 , C12Q1/68 , G01N27/447 , Y10T436/143333
摘要： DNA molecule length can be measured with high precision and efficiency by 1) using such means as electrophoresis gel migration to orient a DNA molecule having a fluorescence label at both its termini into a straight line by its passing through a migration path having in a portion of it an area not more than several micrometers in diameter, detecting the fluorescence label at both the termini at a predetermined location and measuring the interval between the detection of the fluorescence coming from one terminus and that of the fluorescence from the other or by 2) a DNA molecule bound to a fluorescence label at one terminus and to a particle at the other being led as a whole by such means as electric field application into an aperture smaller in diameter than the particle, leaving the particle fixed at the mouth of the aperture to stretch the DNA molecule and detecting the fluorescence position to measure the distance between the bound particle and the bound fluorescence label.
摘要翻译： 可以通过以下方式测量DNA分子长度：1）使用电泳凝胶迁移的方式，通过其经过一部分迁移路径将具有两端的荧光标记的DNA分子定向成直线 它是直径不超过几微米的区域，在预定位置的两个末端检测荧光标记，并测量从一个末端检测荧光和来自另一个末端的荧光检测之间的间隔，或者通过2）a 在一个末端与一个荧光标记结合的DNA分子和另一个的一个粒子整体通过电场施加到比颗粒直径更小的孔隙中，将颗粒固定在孔的口处 拉伸DNA分子并检测荧光位置以测量结合的颗粒和结合的荧光标记之间的距离。
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公开(公告)号：US5824481A
公开(公告)日：1998-10-20
申请号：US812698
申请日：1997-03-06
申请人： Hideki Kambara , Kazunori Okano , Satoshi Takahashi , Keiichi Nagai , Kazuko Kawamoto , Hiroko Furuyama
发明人： Hideki Kambara , Kazunori Okano , Satoshi Takahashi , Keiichi Nagai , Kazuko Kawamoto , Hiroko Furuyama
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6869 , C12Q1/6846
摘要： A DNA analyzing method which bonds a first oligomer of known base sequence to a DNA fragment obtained by digesting a DNA sample with a restrictive enzyme. The oligomer and DNA fragment are hybridized to other oligomers which have the sequences of all combinations of the types of bases within the length of several bases following the known base sequence. The presence or absence of hybridization or complementary DNA strand extension is determined and identifies the DNA fragment terminal sequence from this result. The DNA fragments are then fractionated and analyzed to determine the sequence. This DNA analyzing method provides an effective analysis of mixtures of long DNAs or DNA fragments.
摘要翻译： 将已知碱基序列的第一寡聚体与限制性酶消化DNA样品获得的DNA片段结合的DNA分析方法。 寡聚体和DNA片段与其它寡核苷酸杂交，其具有在已知碱基序列之后的几个碱基长度内的碱基类型的所有组合的序列。 确定是否存在杂交或互补DNA链延伸，并从该结果鉴定DNA片段末端序列。 然后将DNA片段分级并分析以确定序列。 该DNA分析方法提供了长DNA或DNA片段的混合物的有效分析。
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公开(公告)号：US06821734B2
公开(公告)日：2004-11-23
申请号：US10083340
申请日：2002-02-27
申请人： Hideki Kambara , Guohua Zhou , Kazunori Okano , Keiichi Nagai
发明人： Hideki Kambara , Guohua Zhou , Kazunori Okano , Keiichi Nagai
IPC分类号： C12Q168
CPC分类号： C12Q1/682 , C12Q1/6827 , C12Q1/6858 , C12Q2565/537 , C12Q2561/125 , C12Q2531/125 , C12Q2525/301 , C12Q2565/301 , C12Q2565/519
摘要： A method for examining nucleotide sequences of a sample includes adding a group of primers of multiple species to a solution containing the sample and simultaneously synthesizing complementary strands at each of the multiple regions containing the nucleotide sequences; designing the DNA probes with specific sequences elongate the complementary strands by the presence or absence of mutations in the nucleotide sequences, wherein the same number of such DNA probes and the nucleotide sequences are used for complementary strand synthesis; using the nucleotide sequences or their complementary sequences as a template to convert pyrophosphate produced during the elongation reaction to ATP which then reacts with chemiluminescent substrates to develop luminescence to be detected. According to the method, sensitivity is greatly increased by amplification of the amount of pyrophosphate produced in synthesis of the complementary strands without amplifying the copies of nucleotide sequences.
摘要翻译： 检查样品的核苷酸序列的方法包括将一组多种引物添加到含有样品的溶液中，同时在含有该核苷酸序列的多个区域的每个区域合成互补链; 用特定序列设计DNA探针，通过核苷酸序列中存在或不存在突变来延长互补链，其中相同数量的这种DNA探针和核苷酸序列用于互补链合成; 使用核苷酸序列或其互补序列作为模板将在延伸反应期间产生的焦磷酸转化为ATP，然后与化学发光底物反应以产生待检测的发光。 根据该方法，在不扩增核苷酸序列的拷贝的情况下，通过扩增合成互补链中产生的焦磷酸盐的量来大大提高灵敏度。
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公开(公告)号：US5650274A
公开(公告)日：1997-07-22
申请号：US263663
申请日：1994-06-22
申请人： Hideki Kambara , Kazunori Okano , Satoshi Takahashi , Keiichi Nagai , Kazuko Kawamoto , Hiroko Furuyama
发明人： Hideki Kambara , Kazunori Okano , Satoshi Takahashi , Keiichi Nagai , Kazuko Kawamoto , Hiroko Furuyama
CPC分类号： C12Q1/6869 , C12Q1/6846
摘要： A DNA analyzing method comprising ligating the oligomer of a known base sequence to the DNA fragment obtained by digesting a sample with a restriction enzyme, hybridizing an oligomer and DNA fragment using oligomers which have the sequences of all combinations of the types of the bases within the length of several bases following the known base sequence, checking the presence or absence of hybridization or complementary DNA strand extension, identifying the DNA fragment terminal sequence from this result, and fractionating the DNA fragments and analyzing them. This DNA analyzing method provides an effective analysis of mixtures of long DNAs or DNA fragments.
摘要翻译： 一种DNA分析方法，包括将已知碱基序列的寡聚物与通过用限制酶消化样品获得的DNA片段连接，使用寡核苷酸与低聚物和DNA片段杂交，所述寡聚体具有碱基类型的所有组合的序列 根据已知碱基序列的几个碱基的长度，检查杂交或互补DNA链延伸的存在或不存在，从该结果鉴定DNA片段末端序列，并分离DNA片段并分析它们。 该DNA分析方法提供了长DNA或DNA片段的混合物的有效分析。
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公开(公告)号：US5307148A
公开(公告)日：1994-04-26
申请号：US684113
申请日：1991-04-12
IPC分类号： G01N21/64 , G01N27/447
CPC分类号： G01N21/645 , G01N27/44721 , G01N2021/6441
摘要： A multicolor fluorescence detection type electrophoresis apparatus comprises an electrophoretic device having migration lanes in which fragment groups specifically labeled by fluorophores of different light emission peak wavelengths are mixed, added, and migrated. It identifies and detects the fragment groups. It uses laser beams of different wavelengths for exciting the fluorophores in the migration lanes. The laser beams are irradiated to separate positions on the migration lanes for detection of a few kinds of fluorophores at irradiation positions. For the purpose, multicolor optical filters are arranged near the irradiation positions to identify and detect lights emitted from the fluorophores excited by wavelength at every irradiation position.
摘要翻译： 多色荧光检测型电泳装置包括具有迁移通道的电泳装置，其中由不同发光峰波长的荧光团特异性标记的片段组被混合，添加和迁移。 它识别和检测片段组。 它使用不同波长的激光束激发迁移通道中的荧光团。 将激光束照射到迁移道上的分离位置，以便在照射位置检测几种荧光团。 为此，在照射位置附近设置多色滤光片，以识别和检测在每个照射位置由被波长激发的荧光团发出的光。
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公开(公告)号：US5268080A
公开(公告)日：1993-12-07
申请号：US843232
申请日：1992-02-28
申请人： Hideki Kambara , Keiichi Nagai
发明人： Hideki Kambara , Keiichi Nagai
IPC分类号： C12M1/00 , C12M1/34 , C12Q1/68 , G01N21/63 , G01N21/64 , G01N27/447 , G01N33/58 , G01N35/00 , G01N27/26
CPC分类号： G01N27/44782 , G01N27/44721 , G01N2035/00237
摘要： In a DNA detecting method wherein the fluorescence detection type electrophoresis apparatus is used, Texas Red is employed as a fluorophor which labels a DNA fragment, the He-Ne laser having an emission wavelength of 594 nm is used as an excitation light, the excitation light irradiates the line portion of the gel plate simultaneously, and a resulting linear fluorescent image is collected by a cylindrical lens and photodetected, thereby providing an apparatus featuring a higher sensitivity, a smaller size and a lighter weight than conventional apparatuses.
摘要翻译： 在使用荧光检测型电泳装置的DNA检测方法中，使用德克萨斯红作为标记DNA片段的荧光体，将发射波长为594nm的He-Ne激光用作激发光，激发光 同时照射凝胶板的线部分，并通过柱面透镜收集所得的线性荧光图像并进行光电检测，从而提供具有比常规装置更高的灵敏度，更小的尺寸和更轻的重量的装置。
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公开(公告)号：US5192412A
公开(公告)日：1993-03-09
申请号：US800042
申请日：1991-11-29
申请人： Hideki Kambara , Keiichi Nagai
发明人： Hideki Kambara , Keiichi Nagai
IPC分类号： G01N27/447
CPC分类号： G01N27/44721 , G01N27/44782
摘要： An electrophoretic apparatus includes an electrophoresis panel having a fluted glass plate and a non-fluted plane plate. The fluted glass plate is provided with a plurality of substantially parallel narrow electrophoresis grooves and a groove intersecting the electrophoresis grooves for laser beam irradiation. The fluted glass plate and the non-fluted plane plate are closely superposed on each other to form many capillaries adapted to be filled with gel to thereby form many electrophoresis lanes. The electrophoretic apparatus further includes a light measuring instrument to measure fluorescence images at positions which are subjected to laser beam irradiation. Passage of fluorescently labeled sampled fragments through the laser beam irradiation positions in each electrophoresis lane is detected by measuring a change of fluorescence emission with time.
摘要翻译： 电泳装置包括具有槽纹玻璃板和非凹槽平板的电泳面板。 带槽的玻璃板设置有多个基本上平行的窄电泳槽和与用于激光束照射的电泳槽相交的槽。 凹槽玻璃板和非凹槽平面板彼此紧密叠合以形成许多适于填充凝胶的毛细管，从而形成许多电泳通道。 该电泳装置还包括用于在经受激光束照射的位置处测量荧光图像的光测量仪器。 通过测量荧光发射随时间的变化来检测通过每个电泳泳道中的激光束照射位置的荧光标记的采样片段的通过。
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公开(公告)号：US5290419A
公开(公告)日：1994-03-01
申请号：US45135
申请日：1993-04-12
IPC分类号： C12M1/00 , G01N21/64 , G01N27/447 , G01N27/26
CPC分类号： G01N27/44721
摘要： In the multi-color fluorescence detection type electrophoresis apparatus provided with the electrophoresis gel plate, excitation laser source, means to separate the fluorescence images according to each emission wavelength, and the detector of the fluorescence subjected to wavelength selection, two or more laser sources are provided, each of laser lights is irradiated on the sample on a time-sharing basis, and the filter which cuts off the scattered light of each the laser synchronous with the laser beam is installed in front of the wavelength separation means thereby providing simultaneous, quick and real-time analysis of a great number of samples such as DNA and RNA labeled by many types of fluorophores, without overlapping the wavelengths of the excitation light and the fluorescence.
摘要翻译： 在具有电泳凝胶板的多色荧光检测型电泳装置，激发激光源，根据各发光波长分离荧光图像的装置以及进行波长选择的荧光检测器中，两个以上的激光源为 只要分时激光照射在样品上，将与激光同步的每个激光的散射光切断的滤光器安装在波长分离装置的前面，从而提供同时，快速 并对许多类型的荧光团标记的大量样品（如DNA和RNA）进行实时分析，而不会与激发光和荧光的波长重叠。
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公开(公告)号：US4624769A
公开(公告)日：1986-11-25
申请号：US750778
申请日：1985-07-01
申请人： Tamotu Simada , Yoshinori Harada , Hideki Kambara , Keiichi Nagai , Jirou Tokita
发明人： Tamotu Simada , Yoshinori Harada , Hideki Kambara , Keiichi Nagai , Jirou Tokita
CPC分类号： G01N27/44743
摘要： An improved electrophoresis apparatus for nucleic acid fragments in which nucleic acid fragments are separated in the order of their molecular weight in an electrophoresis cell using an electrophoretic gel for determining the base sequence of a nucleic acid, the improvement comprising a nucleic acid fragment specimen supply section for supplying the nucleic acid fragments to the electrophoresis cell, disposed thereover a molecular sieving membrane having a predetermined degree of molecular permeability for separating to remove those high molecular weight ingredients in the nucleic acid fragment specimen other than the fragments intended to be analyzed electrophoretically.
摘要翻译： 一种用于核酸片段的改进的电泳装置，其中核酸片段在电泳细胞中以其分子量的顺序用电泳凝胶分离以确定核酸的碱基序列，其改进包括核酸片段样品供应部分 用于将核酸片段提供给电泳池，将其设置在具有预定分子渗透性分子的分子筛膜上，用于分离以去除除了电泳分析的片段以外的核酸片段样品中的那些高分子量成分。
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