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公开(公告)号：US06867027B1
公开(公告)日：2005-03-15
申请号：US09254344
申请日：1998-07-06
CPC分类号： C12N9/1247 , C12N9/1252
摘要： Disclosed are RNA polymerases consisting of a wild type RNA polymerase provided that at least one of amino acids in the wild type RNA polymerase has been modified to enhance its ability for incorporating 3′-deoxyribonucleotides and derivatives thereof in comparison with the corresponding wild type RNA polymerases. Specifically, disclosed are, for example, the RNA polymerases wherein at least one amino acid present in a nucleotide binding sites of the wild type RNA polymerases such as phenylalanine has been replaced with tyrosine. The RNA polymerases of the present invention are a RNA polymerase which exhibits little or no bias for incorporation between ribonucleotides and 3′-deoxyribonucleotide as well as among ribonucleotides having different base groups and among deoxyribonucleotides having. different base groups.
摘要翻译： 公开了由野生型RNA聚合酶组成的RNA聚合酶，只要野生型RNA聚合酶中的至少一个氨基酸被修饰以增强其与相应的野生型RNA聚合酶相比的3'-脱氧核糖核苷酸及其衍生物的结合能力 。 具体地，例如公开了其中存在于野生型RNA聚合酶的核苷酸结合位点如苯丙氨酸中的至少一个氨基酸已被酪氨酸替代的RNA聚合酶。 本发明的RNA聚合酶是对于核糖核苷酸和3'-脱氧核糖核苷酸之间以及在具有不同碱基的核糖核苷酸和具有不同碱基的脱氧核糖核苷酸之间并入显示很少或没有偏差的RNA聚合酶。 不同的基组。
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2.发明授权公开(公告)号：US08017320B2
公开(公告)日：2011-09-13
申请号：US10554678
申请日：2008-05-14
IPC分类号： C12Q1/68
CPC分类号： G01N33/57446 , C12Q1/6886 , C12Q2600/112 , G01N2333/90203 , G01N2333/988 , G01N2800/56
摘要： Metastatic cancer cells originating from gastric cancer are detected by a method comprising the step of collecting a biological sample from a subject, the step of detecting the presence of at least either aldehyde dehydrogenase or dopa decarboxylase in the biological sample of the subject, and the step of determining that the possibility of containing metastatic cancer cells originating from the gastric cancer in the sample is high when at least either aldehyde dehydrogenase or dopa decarboxylase is present. By the use of these as markers for metastatic cancer cells originating from gastric cancer, the presence or absence of peritoneal metastasis in a gastric cancer patient can be detected rapidly and reliably, and data important for deciding whether intraperitoneal cancer chemotherapy should be applied is provided.
摘要翻译： 通过包括从受试者收集生物样品的步骤的方法检测源自胃癌的转移性癌细胞，检测受试者的生物样品中至少一种醛脱氢酶或多巴脱羧酶的存在的步骤，以及步骤 确定当存在至少一种醛脱氢酶或多巴脱羧酶时，含有样品中源自胃癌的转移性癌细胞的可能性高。 通过使用这些作为来自胃癌的转移性癌细胞的标记物，可以快速可靠地检测胃癌患者腹膜转移的存在或不存在，并且提供了决定是否应用腹膜内癌化疗的重要数据。
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公开(公告)号：US20100035249A1
公开(公告)日：2010-02-11
申请号：US12186009
申请日：2008-08-05
申请人： Yoshihide Hayashizaki , Piero Carninci , Mutsumi Katayama , Shintaro Katayama , Masayoshi Itoh , Matthias Harbers
发明人： Yoshihide Hayashizaki , Piero Carninci , Mutsumi Katayama , Shintaro Katayama , Masayoshi Itoh , Matthias Harbers
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6809 , C12Q1/6834 , C12Q1/6874 , C12Q2535/101 , C12Q2565/537 , C12Q2525/155
摘要： The present invention provides methods for the sequencing of all RNA species within an RNA sample, such as the RNA content obtained from a cell, a tissue, a living organism, or from an artificial source. RNA molecules within the samples are labeled in a RNA-specific manner prior to immobilization on a solid support. One label is used to mark the location of the RNA molecule on the solid support, whereas the second label is used to mark selectively the S′ end of full-length mRNA molecules. RNA molecules are sequenced while being bound to the solid support in one or more sequencing reactions, and sequences of individual RNA molecules can be forwarded to computational analysis for assembling sequence information from individual sequencing reads obtained from the same location on the solid support. Not only unsupervised expression profiling on a genome-wide scale, but also the direct analysis of RNA-RNA interactions become possible as revealed by the analysis of the sequencing information obtained along with genomic information.
摘要翻译： 本发明提供了用于对RNA样品中所有RNA物种进行测序的方法，例如从细胞，组织，活生物体或人造来源获得的RNA含量。 在固定在固体支持物上之前，将样品中的RNA分子以RNA特异性方式标记。 一个标签用于标记RNA分子在固体支持物上的位置，而第二个标记用于选择性标记全长mRNA分子的S'末端。 RNA分子在一个或多个测序反应中与固体支持物结合时进行测序，并且可将单个RNA分子的序列转发到计算分析，以从固体支持物上的相同位置获得的单独测序读数组装序列信息。 不仅在基因组范围内无监督的表达谱，而且通过分析与基因组信息一起获得的测序信息显示，RNA-RNA相互作用的直接分析也是可能的。
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公开(公告)号：US20060160084A1
公开(公告)日：2006-07-20
申请号：US10532975
申请日：2003-10-29
CPC分类号： C12Q1/6844 , C12Q2525/301
摘要： The present invention relates to a process for synthesizing or amplifying efficiently a nucleic acid comprising a target nucleic acid sequence. In the process according to the present invention, a primer comprising in its 3′-end portion a sequence (Ac′) which hybridizes a sequence (A) in the 3′-end portion of the target nucleic acid sequence, and in the 5′-side of said sequence (Ac′) a sequence (B′) which hybridizes the complementary sequence (Bc) of a sequence (B) positioned in the 5′-side of said sequence (A) on the target nucleic acid sequence, wherein {X−(Y−Y′)}/X is in the range of −1.00 to 1.00, in which X denotes the number of bases in said sequence (Ac′), Y denotes the number of bases in the region flanked by said sequences (A) and (B) in the target nucleic acid sequence, and Y′ denotes the number of bases in an intervening sequence between said sequences (Ac′) and (B′) (Y′ may be zero).
摘要翻译： 本发明涉及有效地合成或扩增包含靶核酸序列的核酸的方法。 在本发明的方法中，引物在其3'末端部分包含与目标核酸序列的3'末端部分的序列（A）杂交的序列（Ac'）和5 所述序列（Ac'）的序列（B'）与位于靶核酸序列上的所述序列（A）的5'侧的序列（B）的互补序列（Bc）杂交， 其中{X-（Y-Y'）} / X在-1.00至1.00的范围内，其中X表示所述序列中的碱基数（Ac'），Y表示侧翼的区域中的碱基数 所述序列（A）和（B）在靶核酸序列中，Y'表示所述序列（Ac'）和（B'）之间的间插序列中的碱基数（Y'可以为零）。
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公开(公告)号：US20060113191A1
公开(公告)日：2006-06-01
申请号：US11326368
申请日：2006-01-06
IPC分类号： G01N27/447
CPC分类号： G01N27/44782 , Y10S83/95 , Y10T29/5148 , Y10T29/53391 , Y10T83/0596
摘要： Capillary columns (102) pass through and are inserted in a rubber plate (14), held and fixed by elastic force of rubber, and two-dimensionally arranged on a sample injection side. it fixes the capillary columns (102) arranged on a plane in close contact by holding the same with a holder plate (6a) from below and with a rubber plate (16) from above on a detection side. In order to press the capillary columns (102) against the holder plate 6a and fix the same with the rubber plate (16), a holder plate (6b) fixing the rubber plate (16) to the holder plate (6a) on both sides of the arrangement of the capillary columns (102) is provided.
摘要翻译： 毛细管柱（102）穿过并被插入橡胶板（14）中，橡胶板通过橡胶的弹力保持和固定，并且二维地布置在样品注射侧。 通过将保持板（6a）从下方夹持，并且在检测侧从上方与橡胶板（16）固定布置在平面上的毛细管柱（102）紧密接触。 为了将毛细管柱（102）压靠在保持板6a上并将其固定在橡胶板（16）上，将橡胶板（16）固定到保持板（6a）上的保持板（6b） 在毛细管柱（102）的布置的两侧。
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公开(公告)号：US07049065B2
公开(公告)日：2006-05-23
申请号：US09935592
申请日：2001-08-24
CPC分类号： C12N15/1096
摘要： A method of preparing normalized and/or subtracted cDNA; a method in which the cDNA that is normalized and/or subtracted is in the form of uncloned cDNA (cDNA tester); a method of preparing normalized and/or subtracted cDNA comprising the steps of: (a) preparing cDNA tester; (b) preparing normalization and/or subtraction RNA driver; (c) conducting normalization and/or subtraction in two steps in any order, or conducting normalization/subtraction as a single step and mixing the normalization/subtraction RNA driver with said cDNA tester; (d) adding an enzyme capable of cleaving single strand sites on RNA drivers nonspecifically bound to cDNA tester; (e) removing said single strand RNA driver cleaved in step d) from the tester and removing tester/driver hybrids; and (f) recovering the normalized and/or subtracted cDNA; and a method of efficiently preparing normalized and/or subtracted long-chain, full-coding, and full-length cDNA libraries are provided.
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7.发明申请Method of utilizing the 5'end of transcribed nucleic acid regions for cloning and analysis 审中-公开利用转录核酸区域的5'端克隆和分析的方法公开(公告)号：US20050250100A1
公开(公告)日：2005-11-10
申请号：US10517544
申请日：2003-06-12
CPC分类号： C12N15/1093 , C12N15/1096
摘要： A method is disclosed for obtaining the 5′ends of transcribed regions from a plurality of nucleic acid fragments obtained from biological materials or synthetic pools. DNA fragments encoding the 5′ends are enriched for their individual analysis or for the analysis of concatemers thereof. The sequence information derived from 5′ ends can be used for characterization and cloning of the transcriptome.
摘要翻译： 公开了从生物材料或合成池获得的多个核酸片段获得转录区的5'端的方法。 丰富了编码5'端的DNA片段进行单独分析或分析其并行物。 从5'末端得到的序列信息可用于表达和克隆转录组。
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公开(公告)号：US20050090010A1
公开(公告)日：2005-04-28
申请号：US10469508
申请日：2002-02-25
摘要： The invention discloses a family of cloning vectors capable of cloning nucleic acid inserts of interest of long sizes, with low or reduced background and high efficiency of excision and method for preparing these vectors and library thereof. As example, it is disclosed a cloning vector comprising a construction vector segment (CS) and a replaceable segment (RS), wherein the size of CS is: 36.5 kb≦CS
摘要翻译： 本发明公开了一种克隆载体，其能够克隆具有低或低背景和高效切割的长尺寸感兴趣的核酸插入物及其制备方法及其文库。 例如，公开了包含构建载体片段（CS）和可替换片段（RS）的克隆载体，其中CS的大小为：36.5kb <= CS <38kb，优选CS为37.5kb，包含lox重组 用于网关样重组的Cre重组和/或att重组位点的位点，优选还包括选自以下的背景降低系统：ccdB基因，lox序列，lacZ基因和由限制性内切核酸酶识别的不对称位点序列 。
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公开(公告)号：US06560859B1
公开(公告)日：2003-05-13
申请号：US09402890
申请日：1999-10-14
IPC分类号： G01N2726
CPC分类号： G01N27/44782 , Y10S83/95 , Y10T29/5148 , Y10T29/53391 , Y10T83/0596
摘要： Capillary columns (102) pass through and are inserted in a rubber plate (14), held and fixed by elastic force of rubber, and two-dimensionally arranged on a sample injection side. It fixes the capillary columns (102) arranged on a plane in close contact by holding the same with a holder plate (6a) from below and with a rubber plate (16) from above on a detection side. In order to press the capillary columns (102) against the holder plate 6a and fix the same with the rubber plate (16), a holder plate (6b) fixing the rubber plate (16) to the holder plate (6a) on both sides of the arrangement of the capillary columns (102) is provided.
摘要翻译： 毛细管柱（102）穿过并被插入橡胶板（14）中，橡胶板通过橡胶的弹力保持和固定，并且二维地布置在样品注射侧。 它通过将保持板（6a）从下方夹持而在安装在平面上的毛细管柱（102）上固定，并在检测侧从橡胶板（16）上方固定。 为了将毛细管柱（102）压靠在保持板6a上并将其固定在橡胶板（16）上，将橡胶板（16）固定在两侧的保持板（6a）上的保持板（6b） 提供毛细管柱（102）的布置。
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公开(公告)号：US06294337B1
公开(公告)日：2001-09-25
申请号：US09600747
申请日：2000-08-29
IPC分类号： C12Q168
CPC分类号： C12Q1/6844 , C12Q1/6869 , C12Q2535/101 , C12Q2525/143 , C12Q2521/119 , C12Q2521/101
摘要： A method for sequencing a target DNA fragment in which along with amplification of the target DNA fragment, nucleic acid transcripts are generated using an RNA polymerase and the amplified target DNA fragments are used as templates in the presence of terminators for nucleic acid transcription reaction and the generated nucleic acid transcripts are analyzed, characterized in that the amplification of target DNA fragments and the generation of nucleic acid transcripts are carried out at a constant temperature is disclosed. The amplification of target DNA fragments and the generation of nucleic acid transcripts can be carried out around the room temperature. A DNA sequencing method using a novel method in which without using a thermo-resistant RNA polymerase, the amplification of target DNA fragments and generation of nucleic acid transcript can be carried out simultaneously in parallel is provided.
摘要翻译： 用于对靶DNA片段进行测序的方法，其中连同扩增靶DNA片段，使用RNA聚合酶产生核酸转录物，扩增的靶DNA片段用作核酸转录反应终止子存在下的模板， 分析了产生的核酸转录物，其特征在于公开了靶DNA片段的扩增和核酸转录物的产生在恒温下进行。 靶DNA片段的扩增和核酸转录物的产生可以在室温附近进行。 提供了使用其中不使用耐热RNA聚合酶的新方法的DNA测序方法，靶DNA片段的扩增和核酸转录物的产生可以并行同时进行。
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