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91. 发明专利

JPS6315148A RECOVERY OF TRACE COMPONENT IN GEL 失效
公开(公告)号：JPS6315148A
公开(公告)日：1988-01-22
申请号：JP15781786
申请日：1986-07-07
申请人： HITACHI LTD
发明人： ITO MICHIO , YOSHIDA MOTOKO , OKANO KAZUNOBU
IPC分类号： C07B63/02 , C07B31/00 , C07B63/00 , C07C67/00 , G01N27/26 , G01N27/447
摘要： PURPOSE:To recover an extremely small quantity of a separated component at high purity with high efficiency, by holding a fine gel to the air permeable partition wall in a small chamber and eluting a trace component from the gel using an electrophoretic method to recover the same in a separate small chamber separated by the partition wall. CONSTITUTION:The suction port 14 of a recovery apparatus is pushed against the specific area of a gel containing a component desired to be recovered and a gel piece 1 is sucked onto an air permeable filter through an opening 10. The component contained in the sucked gel piece 1 is taken out from the gel by an electrophoretic method to be recovered. That is, after the gel piece 1 is sucked, the opening part 14 is covered with an ultrafiltration membrane 13 and an aqueous electrolyte solution is introduced into the recovery apparatus from openings 11, 10, 12, 15 to fill the space between a cathode 8 and an anode 9. When a current is made to flow between both electrodes, the component is migrated to be separated in a recovery chamber 3 and can be recovered in a separate container through the opening 10. Ultrafiltration membranes 13, 4 prevent the unrecoverable diffusion of the component due to convection.

92. 发明专利

JPS61114155A Device for manufacturing polyacrylamide gel 失效
标题翻译：制造聚丙烯酰胺凝胶的装置
公开(公告)号：JPS61114155A
公开(公告)日：1986-05-31
申请号：JP23492384
申请日：1984-11-09
申请人： Hitachi Ltd
发明人： YOSHIDA MOTOKO , ITO MICHIO , OKANO KAZUNOBU
IPC分类号： G01N27/26 , B01D57/02 , G01N27/447
CPC分类号： G01N27/44704
摘要： PURPOSE:To manufacture a gel of which the open face in particular is a specular surface and which has excellent sepn. performance by disposing a silicon wafer as a counter plate between glass base plates subjected to a silane coupling agent treatment, injecting a soln. for forming an acrylamide gel between both plates and gelling the soln. CONSTITUTION:The silicon wafer with which the specular surface is obtd. with easy working for surface flattening is used for the counter plate 2. The glass base plates 1, 1 subjected to the surface treatment with the silane coupling agent are combined via the plate 2 and a spacer 3 and are perpendicularly inserted into a vessel 4 for manufacturing the gel. The soln. contg. the acrylamide N-N'-methylene bis-acrylamide for crosslinking and polymn. catalyst is formed in a vessel 6 which can manufacture the soln. to the concn. gradient from a low concn. successively to high concn. Such soln. is successively fed through a peristal pump 5 into the vessel 4. The vessel is so arranged that the gelation progresses after the end of the injection. The gel fixed to the base plates 1 is thus taken out and the plate 2 is stripped away. The open surface has the specular surface in the above-mentioned manner and the good-quality migration image is obtd. without the outflow of albumin, etc. on the surface in the secondary electrophoresis image of serum, etc.
摘要翻译：目的：制造特别是开放面是镜面的凝胶，其具有优异的分辨率。通过在经过硅烷偶联剂处理的玻璃基板之间设置硅晶片作为对置板，注入溶剂来实现。用于在两个板之间形成丙烯酰胺凝胶并使溶胶凝胶化。构成：具有镜面的硅晶片。用于表面平坦化的容易加工用于对置板2.用硅烷偶联剂进行表面处理的玻璃基板1,1经由板2和间隔件3组合，并垂直地插入到容器4中，用于制造凝胶。索恩对比用于交联和聚合的丙烯酰胺N-N'-亚甲基双丙烯酰胺。催化剂形成在可制造溶胶的容器6中。到了从低浓度的梯度先后到高层这样的被连续地通过蠕动泵5进料到容器4中。容器的布置使得凝胶化在注射结束之后进行。因此，固定到基板1上的凝胶被取出并剥离板2。开口表面具有上述方式的镜面，并且质量好的迁移图像是可见的。没有血清等离子电泳图像上的白蛋白等的流出。

93. 发明专利

JPS60236057A TWO-DIMENSIONAL ELECTROPHORESIS DEVICE 失效
公开(公告)号：JPS60236057A
公开(公告)日：1985-11-22
申请号：JP9095784
申请日：1984-05-09
申请人： HITACHI LTD
发明人： ITOU MICHIO , ISHIKAWA KIYOSHI , YOSHIDA MOTOKO , OKANO KAZUNOBU
IPC分类号： G01N27/26 , B01D57/02 , G01N27/447
摘要： PURPOSE:To simplify an operation in the stage of making two-dimensional development by incorporating a one-dimensional gel by rotating motion into a groove provided in a two-dimensional gel and uniting both to one body. CONSTITUTION:A glass plate 1 having a small width is subjected to a silane coupling treatment and a thin layer of a polymer gel 2 consisting of acrylamide is coupled thereto. Such plate is fixed onto a supporting bar 6 and is placed horizontally. Serum which is a sample is injected into the sample hole 3 on the gel. A voltage is impressed to both right and left ends of the gel and electrophoresis is executed so that protein is separated. On the other hand, the concn. gradient gel to be used for the two-dimensions is similarly formed on a glass substrate 5 and a groove 7 for embedding the one-dimensional gel into the gel is provided at the low concn. end of the gel. The one-dimensional gel is transferred into the groove 7 by rotating a driving device 9 around a revolving shaft 8 to turn upward the glass substrate 1 right after the end of the electrophoresis. The space between the one-dimensional gel 2 and the groove 7 is united to one body by low melting agarose and thereafter the voltage is impressed to the gel in the concn. gradient direction, by which the electrophoresis is executed.

94. 发明专利

JPS59169498A Method of stopping staining reactions in separated protein 失效
标题翻译：在分离蛋白中停止染色反应的方法
公开(公告)号：JPS59169498A
公开(公告)日：1984-09-25
申请号：JP4424783
申请日：1983-03-18
申请人： Hitachi Ltd
发明人： OKANO KAZUNOBU , ITOU MICHIO , YOSHIDA MOTOKO
IPC分类号： G01N33/68 , C12Q1/30
摘要： PURPOSE: An electrophoretically separated protein is activated with peroxidase to develop its color with hydrogen peroxide, then the excessive hydrogen peroxide is decomposed with a catalase to stop staining whereby protein spots are detected in stabilized states.
CONSTITUTION: Proteins on a support are subjected to electrophoresis to separate into individuals, then dipped in, as such, in a hemin solution so that the proteins absorb hemin and become active to peroxidase. Then, the support is dipped in a solution of pigment precursor and the protein spots are stained with hydrogen peroxide. Then, the excessive amount of hydrogen peroxide is decomposed with catalase to stop staining on the protein spots.
USE: Inspections of medical specimens.
COPYRIGHT: (C)1984,JPO&Japio
摘要翻译：目的：电泳分离的蛋白质用过氧化物酶活化，以过氧化氢显色，然后用过氧化氢酶分解过量的过氧化氢，停止染色，从而在稳定状态下检测蛋白质斑点。构成：将载体上的蛋白质进行电泳分离成个体，然后浸入血红素溶液中，使得蛋白质吸收血红素并变得对过氧化物酶具有活性。然后，将载体浸渍在颜料前体溶液中，蛋白质斑点用过氧化氢染色。然后，用过氧化氢酶分解过量的过氧化氢，停止对蛋白质斑点的染色。用途：检查医用标本。

95. 发明专利

JP2003159076A PRIMER FOR PCR AND METHOD FOR DETERMINING BASE SEQUENCE OF PRIMER FOR PCR 审中-公开
公开(公告)号：JP2003159076A
公开(公告)日：2003-06-03
申请号：JP2001360321
申请日：2001-11-27
申请人： HITACHI LTD
发明人： UEMATSU SENSHU , OKANO KAZUNOBU , IRIE TAKASHI
IPC分类号： G01N27/447 , C12N15/09 , C12Q1/68
摘要： PROBLEM TO BE SOLVED: To provide a PCR primer which causes the addition of adenine to a DNA fragment with a DNA polymerase on PCR but gives a single peak without changing a condition for the adenine addition reaction. SOLUTION: The PCR is carried out by preparing four PCR primers each having an anchor sequence at the 5'-terminal and having one of A, C, G and T bases at the 5'-terminal of the anchor sequence, determining the adenine addition efficiency of each primer in each PCR, screening anchor sequences causing adenine addition to determine the anchor sequence easily causing the adenine addition, and using the primer having the determined anchor sequence. COPYRIGHT: (C)2003,JPO

96. 发明专利

JP2003135097A METHOD FOR TESTING NUCLEIC ACID SEQUENCE 审中-公开
公开(公告)号：JP2003135097A
公开(公告)日：2003-05-13
申请号：JP2001331853
申请日：2001-10-30
申请人： HITACHI LTD
发明人： KANBARA HIDEKI , SHU KOKUKA , OKANO KAZUNOBU , NAGAI KEIICHI
IPC分类号： C12N15/09 , C12Q1/68
摘要： PROBLEM TO BE SOLVED: To provide an efficient method for testing nucleic acid sequences in a sample having many measuring sites based on chemiluminescence. SOLUTION: The method comprises a process in which a primer group consisting of two or more kinds of primers is put into the sample, and complementary chain syntheses are performed simultaneously for all regions containing the targeting nucleic acid sequences, respectively, a process in which complementary chain extensions initiated with each of DNA probes of a DNA group and the corresponding target, are carried out. The DNA group contains the DNA probes having sequences, the extension of whose complementary chain is influenced by the existence of the mutation in the target nucleic acid sequence, of the same number as the kinds of targets, and a process in which complementary chain extension reactions using targets or complementary sequences of the targets as templates, a reaction for converting the pyrophosphoric acid formed by the complementary chain extension reactions into ATP and a reaction of the ATP with a chemiluminescent substrate to generate light emission are carried out in subcells of a reaction vessel which are separated for each target. And, a process in which mutations in the targets are detected by detecting the light emissions. The detection sensitivity is increased by amplifying the amount of the pyrophosphoric acid formed by the complementary chain synthesis without amplifying the number of copies of a target.

97. 发明专利

JP2000106900A ANALYSIS OF MIXED DNA FRAGMENTS 失效
公开(公告)号：JP2000106900A
公开(公告)日：2000-04-18
申请号：JP28628698
申请日：1998-10-08
申请人： HITACHI LTD
发明人： UEMATSU SENSHU , KANBARA HIDEKI , OKANO KAZUNOBU
IPC分类号： C12N15/09 , C12Q1/68 , G01N33/53 , G01N33/50 , G01N33/566
摘要： PROBLEM TO BE SOLVED: To provide a method for analyzing mixed DNA fragments which enables us to perform a gene expression monitoring method. SOLUTION: This method for analyzing mixed DNA fragments comprises the following processes and enables us to compare plural specimens without being affected by reproducibility of PCR: (1) individually combining oligomers 11 and 12 capable of complementarily combining with primers 21 and 22 mutually having the same dissolution temperature and the same length with DNA fragments 3 and 4 in each group of DNA fragment groups; (2) mixing DNA fragments 5 and 6 of each group in which oligomers are combined; (3) simultaneously subjecting the DNA fragments 5 and 6 of each group in which oligomers are combined to PCR with primers 21 and 22 respectively complementary to oligomers 11 and 12 and corresponding to each group in an identical vessel 29; (4) detecting DNA fragments multiplied through PCR by an electrophoretic separation procedure.

98. 发明专利

JPH09154599A DETECTION OF DNA 失效
公开(公告)号：JPH09154599A
公开(公告)日：1997-06-17
申请号：JP31937795
申请日：1995-12-07
申请人： HITACHI LTD
发明人： KANBARA HIDEKI , OKANO KAZUNOBU
IPC分类号： G01N33/50 , C12N15/09 , C12Q1/68 , G01N30/88 , G01N33/53 , G01N33/566
摘要： PROBLEM TO BE SOLVED: To simply and highly sensitively detect a specific DNA by hybridizing the target DNA with a primer, synthesizing a complementary chain containing a fluorescent light label, cleaving the chain, separating the produced oligomers and subsequently detecting the fluorescent light label. SOLUTION: This method for detecting the DNA comprises hybridizing a target DNA 1 with a primer 2 comprising an oligomer, synthesizing a complementary chain 3 containing at least a fluorescent light-labeled deoxynucleotide triphosphate and deoxynucieotlde α-thiotriphosphate as substrates on the primer, cleaving the chain of the DNA produced by the synthesis of the complementary chain by a chemical reaction method or by an enzymatic reaction using an endonuclease, etc., to produce one or more kinds of oligomers 6, separating the produced fragments by a gel electrophoresis method, a liquid chromatograph method, etc., and subsequently measuring the fluorescent light generated from the fluorescent light-labeled fragment 4. Thereby, the specific target DNA can highly sensitively be detected without applying a temperature cycle.

99. 发明专利

JPH0965880A DNA ANALYSIS 失效
公开(公告)号：JPH0965880A
公开(公告)日：1997-03-11
申请号：JP22074495
申请日：1995-08-29
申请人： HITACHI LTD
发明人： KANBARA HIDEKI , OKANO KAZUNOBU
IPC分类号： G01N33/50 , C12N15/09 , C12Q1/68
摘要： PROBLEM TO BE SOLVED: To obtain a new primer as a primer DNA for a DNA complementary strand synthesis reaction, comprising a part to which a complementary strand is bonded, constituting a 3' projected end at the 3' end of the DNA primer, capable of analyzing a DNA sequence by a simple operation. SOLUTION: This new primer is a primer DNA for a DNA complementary strand synthesis reaction, comprises a part to which a complementary strand is bonded, constitutes a 3' projected end at the 3' end of the DNA primer and is composed of a double strand DNA oligomer. The primer has a 3' end projected part to be hybridized with a restriction enzyme scission part and is useful for a DNA analysis capable of determining a DNA sequence requiring a trouble by a simple operation. The primer is obtained by forming a double strand DNA composed of complementary strands 22 and 23 at a part of the DNA oligomer, bonding a 3' projected end constituted of a restriction enzyme recognition part 24 and a selection part 25 to its 3' end or bonding the 3' projected ends 24 and 25 to the 3' end of a DNA chain having a self-hybridizing part 26.

100. 发明专利

JPH0739399A METHOD FOR ANALYZING DNA 失效
公开(公告)号：JPH0739399A
公开(公告)日：1995-02-10
申请号：JP18962493
申请日：1993-07-30
申请人： HITACHI LTD
发明人： KANBARA HIDEKI , OKANO KAZUNOBU , TAKAHASHI SATOSHI , NAGAI KEIICHI
IPC分类号： C12N15/09 , C12Q1/68 , G01N33/53 , G01N37/00
摘要： PURPOSE:To efficiently carry out a method for analyzing a DNA useful for determining, etc., the base sequence of the DNA in which processes for cleaving a DNA chain, discriminating and fractionating the resultant DNA fragments and determining the base sequence are provided and DNA fragments having many different sequences can simultaneously be analyzed. CONSTITUTION:This method for analyzing a DNA comprises cleaving a DNA sample 1 with a restriction enzyme, connecting an oligomer having a known sequence to the terminal of the resultant DNA fragments, inducing the hybridization between oligomers 3 having the sequence of all the combinations of base species within the range of several bases following the known sequence and the DNA fragments using a DNA probe chip 4 having the oligomers 3 immobilized on a solid surface, detecting the presence or absence of the hybridization, knowing the terminal sequences of the resultant DNA fragments from the detected results and fractionating the obtained DNA fragments or directly analyze the DNA fragments.

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