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1. 发明公开

EP0989193A2 DNA analyzing method 失效
标题翻译： DNS分析Verfahren
公开(公告)号：EP0989193A2
公开(公告)日：2000-03-29
申请号：EP99124555.6
申请日：1994-06-23
申请人： Hitachi, Ltd.
发明人： Kambara, Hideki , Okano, Kazunori , Takahashi, Satoshi , Nagai, Keiichi , Kawamoto, Kazuko , Furuyama, Hiroko
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6869 , C12Q1/6846 , C12Q2525/191 , C12Q2523/107 , C12Q2521/319 , C12Q2537/143
摘要： A DNA analyzing method which bonds the oligomer of the known base sequence to the DNA fragment obtained by digesting sample 1 with restrictive enzyme, causes hybridization between oligomer 3 and DNA fragment using the oligomers 3 which have the sequences of all combinations of the types of the bases within the length of several bases following the known base sequence, checks presence or absence of the hybridization or complementary DNA strand extension, identifies the DNA fragment terminal sequence from this result, and fractionates the DNA fragments and analyzes them or analyzes them as they are. This DNA analyzing method provides an effective analysis of mixtures of long DNAs or DNA fragments.
摘要翻译：将已知碱基序列的寡聚物与通过用限制性酶消化样品1获得的DNA片段结合的DNA分析方法，使用具有所有类型的所有组合的序列的低聚物3和DNA片段之间的杂交，在已知碱基序列之后的几个碱基长度内的碱基检查杂交或互补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，因为它们是。该DNA分析方法提供了长DNA或DNA片段的混合物的有效分析。

2. 发明公开

EP0959141A2 Method of preparing nucleic acid sample for rare expressed genes and analysing method using the nucleic acid prepared thereby 审中-公开
标题翻译：一种用于纯化的罕见表达的基因和分析程序的核酸的方法，以便
公开(公告)号：EP0959141A2
公开(公告)日：1999-11-24
申请号：EP99109795.7
申请日：1999-05-18
申请人： Hitachi, Ltd.
发明人： Muramatsu, Takamichi , Fujita, Takeshi , Kiyama, Masaharu , Irie, Takashi , Okano, Kazunori
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6806 , C12Q1/6834
摘要： Focusing that abundant class genes abundantly present in a nucleic acid sample can comparatively easily be analyzed and readily be removed in a selective manner, a nucleic acid sample for expression analysis of rare expressed genes can be obtained by removing abundant genes therefrom, and analyses based upon the sample can be achieved.
摘要翻译：聚焦并丰富类基因中大量存在的核酸样品中可以比较容易地进行分析，并容易地以选择性的方式被去除，为的稀有过表达的基因的表达分析的核酸样品可以通过从除去丰富基因那里获得，并且分析基于样品就可以实现。

3. 发明公开

EP0630972A2 DNA analyzing method 失效
标题翻译： DNS分析Verfahren。
公开(公告)号：EP0630972A2
公开(公告)日：1994-12-28
申请号：EP94109745.3
申请日：1994-06-23
申请人： HITACHI, LTD.
发明人： Kambara, Hideki , Okano, Kazunori , Takahashi, Satoshi , Nagai, Keiichi , Kawamoto, Kazuko, Nr. 309, Hitachi Ozaki-haitsu , Furuyama, Hiroko, Nr. 110, Hitachi Ozaki-haitsu
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6869 , C12Q1/6846 , C12Q2525/191 , C12Q2523/107 , C12Q2521/319 , C12Q2537/143
摘要： A DNA analyzing method which bonds the oligomer of the known base sequence to the DNA fragment obtained by digesting sample 1 with restrictive enzyme, causes hybridization between oligomer 3 and DNA fragment using the oligomers 3 which have the sequences of all combinations of the types of the bases within the length of several bases following the known base sequence, checks presence or absence of the hybridization or complementary DNA strand extension, identifies the DNA fragment terminal sequence from this result, and fractionates the DNA fragments and analyzes them or analyzes them as they are. This DNA analyzing method provides an effective analysis of mixtures of long DNAs or DNA fragments.
摘要翻译：将已知碱基序列的寡聚物与通过用限制性酶消化样品1获得的DNA片段结合的DNA分析方法，使用具有所有类型的所有组合的序列的低聚物3和DNA片段之间的杂交，在已知碱基序列之后的几个碱基长度内的碱基检查杂交或互补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，因为它们是。该DNA分析方法提供了长DNA或DNA片段的混合物的有效分析。

4. 发明授权

EP0649852B1 Fractionation method for nucleotide fragments 失效
标题翻译：核苷酸片段的Fraktioniermethode
公开(公告)号：EP0649852B1
公开(公告)日：2000-01-05
申请号：EP94116671.2
申请日：1994-10-21
申请人： HITACHI, LTD.
发明人： Kambara, Hideki , Okano, Kazunori , Uematsu, Chihiro
IPC分类号： C07H1/06 , C12Q1/68
CPC分类号： G01N27/44743 , C12Q1/6837 , C12Q1/6874 , G01N27/447 , C12Q2535/101 , C12Q2525/179 , C12Q2565/537 , C12Q2525/191

5. 发明公开

EP0767240A3 DNA sequencing method and DNA sample preparation method 失效
标题翻译： DNA测序及其制备方法
公开(公告)号：EP0767240A3
公开(公告)日：1999-12-22
申请号：EP96114907.7
申请日：1996-09-17
申请人： HITACHI, LTD.
发明人： Kambara, Hideki , Okano, Kazunori
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6869 , C12N15/1096 , C12Q2535/101 , C12Q2525/191 , C12Q2521/301
摘要： The DNA sequencing of the present invention comprises:
(1) a step of digesting a sample DNA with a restriction enzyme to obtain a DNA fragment; (2) a step of introducing an oligonucleotide having a definite base sequence into the DNA fragment at the 3' terminus thereof; (3) a step of performing a complementary strand extension reaction, using a labeled primer which comprises a first base sequence portion complementary to at least a part of the base sequence of the oligonucleotide, a second base sequence portion complementary to the recognition base sequence recognized by the restriction enzyme and a third base sequence of a possible combination of 1 to 4 bases, by using, as a template, a single strand of the DNA fragment having the oligonucleotide introduced thereinto to obtain a labeled extended primer having a base sequence complementary to the single strand of the DNA fragment; (4) a step of proceeding:
(a) a step of performing a sequencing reaction using the labeled primer by using, as a template, the single strand of the DNA fragment having the oligonucleotide introduced thereinto, and, (b) a step of performing a sequencing reaction using the extended labeled primer by using, as a template, a part of the single strand of the sample DNA having the base sequence of the single strand of the DNA fragment and a contiguous sequence adjacent thereto, or the single strand of the sample DNA; and, (5) a step of subjecting products of the sequencing reaction to electrophoresis to determine the base sequence of the DNA fragment and at least a part of the base sequence of the sample DNA adjacent to the base sequence of the DNA fragment.

6. 发明公开

EP0767240A2 DNA sequencing method and DNA sample preparation method 失效
标题翻译： DNS Sequenzierungs- und Vorbereitungsverfahren
公开(公告)号：EP0767240A2
公开(公告)日：1997-04-09
申请号：EP96114907.7
申请日：1996-09-17
申请人： HITACHI, LTD.
发明人： Kambara, Hideki , Okano, Kazunori
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6869 , C12N15/1096 , C12Q2535/101 , C12Q2525/191 , C12Q2521/301
摘要： The DNA sequencing of the present invention comprises:

(1) a step of digesting a sample DNA with a restriction enzyme to obtain a DNA fragment;
(2) a step of introducing an oligonucleotide having a definite base sequence into the DNA fragment at the 3' terminus thereof;
(3) a step of performing a complementary strand extension reaction, using a labeled primer which comprises a first base sequence portion complementary to at least a part of the base sequence of the oligonucleotide, a second base sequence portion complementary to the recognition base sequence recognized by the restriction enzyme and a third base sequence of a possible combination of 1 to 4 bases, by using, as a template, a single strand of the DNA fragment having the oligonucleotide introduced thereinto to obtain a labeled extended primer having a base sequence complementary to the single strand of the DNA fragment;
(4) a step of proceeding:

(a) a step of performing a sequencing reaction using the labeled primer by using, as a template, the single strand of the DNA fragment having the oligonucleotide introduced thereinto, and,
(b) a step of performing a sequencing reaction using the extended labeled primer by using, as a template, a part of the single strand of the sample DNA having the base sequence of the single strand of the DNA fragment and a contiguous sequence adjacent thereto, or the single strand of the sample DNA; and,

(5) a step of subjecting products of the sequencing reaction to electrophoresis to determine the base sequence of the DNA fragment and at least a part of the base sequence of the sample DNA adjacent to the base sequence of the DNA fragment.
摘要翻译：本发明的DNA测序包括：（1）用限制酶消化样品DNA以获得DNA片段的步骤; （2）将具有确定碱基序列的寡核苷酸引入其3'末端的DNA片段的步骤; （3）使用包含与寡核苷酸的碱基序列的至少一部分互补的第一碱基序列部分的标记引物进行互补链延伸反应的步骤，与识别的识别碱基序列互补的第二碱基序列部分通过限制性内切酶和1〜4个碱基的可能组合的第3碱基序列，通过使用引入寡核苷酸的DNA片段的单链作为模板，得到具有与 DNA片段的单链; （4）进行以下步骤：（a）通过使用引入了寡核苷酸的DNA片段的单链作为模板，使用标记引物进行测序反应的步骤，（b）步骤使用延伸的标记引物进行测序反应，通过使用具有DNA片段的单链的碱基序列和与其相邻的连续序列的样品DNA的单链的一部分作为模板，或单链的样品DNA; 和（5）使测序反应产物进行电泳以确定DNA片段的碱基序列和与DNA片段的碱基序列相邻的样品DNA的至少一部分碱基序列的步骤。

7. 发明公开

EP0578138A2 Gene-detecting method 失效
标题翻译：基因检测方法
公开(公告)号：EP0578138A2
公开(公告)日：1994-01-12
申请号：EP93110550.6
申请日：1993-07-01
申请人： HITACHI, LTD.
发明人： Kambara, Hideki , Okano, Kazunori , Murakawa, Katsuji
IPC分类号： C12Q1/68
CPC分类号： C12Q1/682 , C12Q1/6823 , C12Q2537/157 , C12Q2521/319 , C12Q2537/155
摘要： A fluorescence-labelled DNA probe 11 is hybridized with a specimen 7 to form 9 double-stranded hybrid 8; successive decomposition and separation of the hybridized DNA probe with the aid of an enzyme specifically decomposing the double-stranded hybrid 8 are repeated to high speed-decompose the DNA probe alone hybridized to the specimen DNA; and after a shortened DNA probe is amplification-produced in this way, the shortened DNA probe 20 and unreacted DNA probe 21 are separately discriminated and detected to detect the specimen DNA 7 with high sensitivity. The shortened DNA probe can be produced in amounts which are larger by several figures than the amount of the specimen DNA in several minutes and without raising and lowering the reaction temperature, so that the specimen DNA can be detected rapidly.
摘要翻译：荧光标记的DNA探针11与样本7杂交以形成9个双链杂合体8; 重复高速分析杂交DNA探针，借助于特异性分解双链杂交物8的酶，高速分解单独的与探针DNA杂交的DNA探针; 用这种方法扩增产生缩短的DNA探针后，分别缩短DNA探针20和未反应的DNA探针21并进行检测，以高灵敏度检测样本DNA 7。缩短了的DNA探针的生成量可以比试样DNA的数分钟大几个数量级，而且不会升高和降低反应温度，因此可以快速检测样本DNA。

8. 发明授权

EP0989193B1 DNA analyzing method 失效
标题翻译： DNA分析方法
公开(公告)号：EP0989193B1
公开(公告)日：2002-09-04
申请号：EP99124555.6
申请日：1994-06-23
申请人： Hitachi, Ltd.
发明人： Kambara, Hideki , Okano, Kazunori , Takahashi, Satoshi , Nagai, Keiichi , Takakaski, Kazuko , Hirabayashi, Hiroko
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6869 , C12Q1/6846 , C12Q2525/191 , C12Q2523/107 , C12Q2521/319 , C12Q2537/143

9. 发明公开

EP0718409A3 DNA preparation method 失效
标题翻译：制备工艺DNS
公开(公告)号：EP0718409A3
公开(公告)日：1999-02-03
申请号：EP95120038.5
申请日：1995-12-19
申请人： HITACHI, LTD.
发明人： Kambara, Hideki , Okano, Kazunori , Fujimori, Kiyoshi
IPC分类号： C12Q1/68 , C12N15/10
CPC分类号： C12Q1/6855
摘要： A DNA preparation method is disclosed, said method comprising a first step of producing a plurality of DNA fragments having different lengths by digestion of double strand DNA; a second step of introducing into said DNA fragments the oligomer having a known base sequence at the portion digested in the first step by ligation; and a third step of carrying out complementary strand extension of the DNA fragments with primer having selective sequence on its 3'-terminus obtained in said second step and increasing the number of DNA fragment copies.

10. 发明公开

EP0582256A3 Polynucleotide detecting method and apparatus 失效
标题翻译：多核苷酸检测方法和装置
公开(公告)号：EP0582256A3
公开(公告)日：1997-09-17
申请号：EP93112368.1
申请日：1993-08-02
申请人： HITACHI, LTD.
发明人： Okano, Kazunori , Kambara, Hideki , Nagai, Keiichi
IPC分类号： C12Q1/68 , B01D57/02 , G01N27/447
CPC分类号： C12Q1/6823 , C12Q1/68 , C12Q1/6813 , G01N27/44721
摘要： Fluorescent-labeled probes 2 hybridizable with target polynucleotide chain 1 are used as fluorescent-labeled probes, and both are reacted to obtain a fluorescent-labeled probe 3 bound to the target polynucleotide 1. After electrophoretically separating the unreacted excess fluorescent-labeled probes, electrophoresis is further continued at a temperature above the denaturing temperature of the complex 3 of target polynucleotide and fluorescent-labeled probe to release the fluorescent-labeled probe 4 which has been bound to the target polynucleotide. Laser beam 6 is applied to the released fluorescent-labeled probe 4 and the generated fluorescence is detected by fluorescence detector 7. Thereby, the present invention enables successive automatic prosecution of a series of operating step and provides a high-sensitivity and high-precision quantitative polynucleotide detecting method suited for application to a high-throughput automated detecting apparatus.

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