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1. 发明授权

US06183998B2 Method for reversible modification of thermostable enzymes 有权
标题翻译：热稳定酶可逆修饰的方法
公开(公告)号：US06183998B2
公开(公告)日：2001-02-06
申请号：US09183950
申请日：1998-10-31
申请人： Igor Ivanov , Dirk Löffert , Jie Kang , Joachim Ribbe , Kerstin Steinert
发明人： Igor Ivanov , Dirk Löffert , Jie Kang , Joachim Ribbe , Kerstin Steinert
IPC分类号： C12P1934
CPC分类号： C12N9/99 , C12N9/1252 , C12Q1/6848 , C12Q2527/125 , C12Q2527/101
摘要： A method for the amplification of a target nucleic acid is disclosed comprising the steps of reacting a nucleic acid with an amplification reaction mixture and a modified thermostable enzyme, wherein said modified thermostable polymerase is prepared by a reaction of a mixture of a thermostable polymerase and a chemical modifying reagent. The chemical modification reagent is an aldehyde, preferably formaldehyde. Essentially complete inactivation of the enzyme at ambient temperatures is achieved, with recovery of enzymatic activity at temperatures above 50° C.
摘要翻译：公开了扩增靶核酸的方法，其包括使核酸与扩增反应混合物和修饰的热稳定酶反应的步骤，其中所述修饰的热稳定聚合酶通过热稳定聚合酶和化学改性剂。化学改性剂是醛，优选甲醛。实现了在环境温度下酶的基本完全失活，并且在高于50℃的温度下回收酶活性。

2. 发明授权

US08124391B2 Thermostable polymerases from Thermococcus pacificus 失效
标题翻译：热太阳能热稳定聚合酶
公开(公告)号：US08124391B2
公开(公告)日：2012-02-28
申请号：US10398011
申请日：2001-10-05
申请人： Katja Decker , Dirk Löffert , Jie Kang
发明人： Katja Decker , Dirk Löffert , Jie Kang
IPC分类号： C12N9/12
CPC分类号： C12N9/1252
摘要： The invention relates to a thermostable polymerase based on thermococcus pacificus; DNA molecules which code for one such polymerase; expression vectors; host cells; methods for producing one such polymerase and the use thereof for polymerising nucleic acid, especially in the polymerase chain reaction.
摘要翻译：本发明涉及一种基于热电球菌的热稳定聚合酶; 编码一种这样的聚合酶的DNA分子; 表达载体宿主细胞; 用于生产一种这样的聚合酶的方法及其用于聚合核酸的用途，特别是在聚合酶链式反应中。

3. 发明授权

US06300069B1 Generation and amplification of nucleic acids from ribonucleic acids 有权
标题翻译：来自核糖核酸的核酸的产生和扩增
公开(公告)号：US06300069B1
公开(公告)日：2001-10-09
申请号：US09304452
申请日：1999-05-03
申请人： Andreas Missel , Dirk Löffert , Jie Kang , Christian Korfhage
发明人： Andreas Missel , Dirk Löffert , Jie Kang , Christian Korfhage
IPC分类号： C12Q168
CPC分类号： C12N15/1096 , C12Q1/686 , C12Q2527/125 , C12Q2521/107
摘要： Novel compositions and methods useful for the generation of nucleic acids from a ribonucleic acid template and further nucleic acid replication are disclosed. It is shown that the generation and amplification of nucleic acids by methods that utilize two or more different polymerases, such as reverse transcriptase-polymerase chain reaction (RT-PCR), are dramatically more sensitive and efficient in the presence of a homopolymeric nucleic acid. Homopolymeric nucleic acids have been found to reduce or negate the inhibitory effect reverse transcriptases have on DNA polymerase activity. It is demonstrated that this inhibition-relieving effect of homopolymeric nucleic acids is general in nature; independent of the chemical species of homopolymer used, or the chemical composition of the polymerization reaction mixture.
摘要翻译：公开了用于从核糖核酸模板产生核酸和进一步核酸复制的新型组合物和方法。显示通过利用两种或多种不同聚合酶的方法（如逆转录酶 - 聚合酶链反应（RT-PCR））在均聚核酸存在下显着更灵敏和有效地产生和扩增核酸。已经发现均聚核酸减少或否定逆转录酶对DNA聚合酶活性的抑制作用。证明均聚核酸的这种抑制作用是一般性的; 独立于所用均聚物的化学物质，或聚合反应混合物的化学组成。

4. 发明授权

US08637288B2 Thermostable chimeric nucleic acid polymerases and uses thereof 失效
标题翻译：热稳定嵌合核酸聚合酶及其用途
公开(公告)号：US08637288B2
公开(公告)日：2014-01-28
申请号：US10216682
申请日：2002-08-08
申请人： Dirk Löffert , Andreas Missel , Jie Kang
发明人： Dirk Löffert , Andreas Missel , Jie Kang
IPC分类号： C12N9/12
CPC分类号： C12P19/34 , C07K2319/00 , C12N9/1252
摘要： Novel thermostable chimeric nucleic acid polymerases and methods for their generation and use are disclosed. It is shown that these chimeric nucleic acid polymerases, such as DNA polymerases, can be constructed using enzymatically active domains, isolated from different proteins or chemically synthesized. It is demonstrated that chimeric nucleic acid polymerases of the present invention possess the chemical and physical properties of their component domains (e.g., exonuclease activity, thermostability) and that the chimeric polymerases are thermostable.
摘要翻译：公开了新型热稳定嵌合核酸聚合酶及其产生和使用的方法。显示这些嵌合核酸聚合酶，例如DNA聚合酶，可以使用从不同蛋白质分离的化学合成的酶活性结构域构建。证明本发明的嵌合核酸聚合酶具有其组分结构域的化学和物理性质（例如外切核酸酶活性，热稳定性），并且嵌合聚合酶是热稳定的。

5. 发明申请

US20110124050A1 METHOD FOR SYNTHESIZING A CDNA IN A SAMPLE IN AN ENZYMATIC REACTION 审中-公开
标题翻译：用于在酶反应中合成样品中的CDNA的方法
公开(公告)号：US20110124050A1
公开(公告)日：2011-05-26
申请号：US12673251
申请日：2008-02-13
申请人： Holger Engel , Subrahmanyam Yerramilli , Martin Kreutz , Dirk Löffert , Christian Korfhage
发明人： Holger Engel , Subrahmanyam Yerramilli , Martin Kreutz , Dirk Löffert , Christian Korfhage
IPC分类号： C12P19/34 , C12N9/12
CPC分类号： C12Q1/6806 , C12Q1/6848 , C12Q2527/101 , C12Q2521/131 , C12Q2521/107
摘要： The present invention relates to a method for synthesizing a cDNA in a sample in an enzymatic reaction, characterized in that the method comprises the steps: simultaneously providing of a first enzyme with polyadenylation activity, a second enzyme with reverse transcriptase activity, a buffer, at least one ribonucleotide, at least one deoxyribonucleotide, an anchor oligonucleotide, adding of a sample comprising a ribonucleic acid and incubating the agents from the preceding steps in one or more temperature steps, which are selected so that the first enzyme and the second enzyme display activity, characterized in that additionally an amplification takes place in the same reaction mixture. The invention relates further to a reaction mixture comprising a first enzyme with polyadenylation activity, a second enzyme with reverse transcriptase activity, optionally a buffer, optionally at least one ribonucleotide, optionally at least one deoxyribonucleotide, optionally an anchor oligonucleotide and an enzyme with DNA synthesis activity.
摘要翻译：本发明涉及一种在酶促反应中合成样品中的cDNA的方法，其特征在于该方法包括以下步骤：同时提供具有多聚腺苷酸化活性的第一种酶，具有逆转录酶活性的第二种酶，缓冲液，至少一个核糖核苷酸，至少一个脱氧核糖核苷酸，锚定寡核苷酸，加入包含核糖核酸的样品，并在一个或多个温度步骤中孵育来自前述步骤的试剂，其被选择为使得第一酶和第二酶显示活性其特征在于另外在相同的反应混合物中进行扩增。本发明进一步涉及包含具有多聚腺苷酸化活性的第一酶的反应混合物，具有逆转录酶活性的第二种酶，任选的缓冲液，任选的至少一种核糖核苷酸，任选的至少一种脱氧核糖核苷酸，任选的锚定寡核苷酸和具有DNA合成的酶活动。

6. 发明授权

US09683255B2 Method for activating a nucleic acid for a polymerase reaction 有权
公开(公告)号：US09683255B2
公开(公告)日：2017-06-20
申请号：US11991435
申请日：2006-09-11
申请人： Christian Korfhage , Dirk Löffert
发明人： Christian Korfhage , Dirk Löffert
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6806 , C12Q1/6844 , C12Q2527/107
摘要： The present invention concerns a method for activating a nucleic acid for a polymerase reaction with the steps: (a) Heating a nucleic acid to a temperature of 55° C. to 80° C., (b) cooling the nucleic acid to a temperature at which a polymerase shows no substantial decrease in activity, and (c) starting the polymerase reaction by the addition of a heat-labile polymerase to the nucleic acid.

7. 发明授权

US09587267B2 Quantification of nucleic acids 有权
标题翻译：核酸定量
公开(公告)号：US09587267B2
公开(公告)日：2017-03-07
申请号：US13255696
申请日：2010-03-09
申请人： Nan Fang , Andreas Missel , Dirk Löffert
发明人： Nan Fang , Andreas Missel , Dirk Löffert
IPC分类号： C12P19/34 , C12Q1/68 , C07H21/02 , C07H21/04 , C07H21/00
CPC分类号： C12Q1/6806 , C12Q2521/325
摘要： The invention relates to a method for the quantification of one or more nucleic acids in a sample, for example: making a sample available which contains at least one nucleic acid to be quantified, adding an oligonucleotide probe, the oligonucleotide probe comprising a sequence which can specifically hybridize to the nucleic acid to be quantified or to a common sequence of the nucleic acids to be quantified, incubating the sample under conditions which allow the hybridization of the oligonucleotide probe to the nucleic acid(s) to be quantified, incubating the sample under conditions which allow the extension of hybridized probes, the nucleic acid(s) serving as a template in each case, removing the non-hybridized probes from the sample and quantifying the hybridized oligonucleotide probes to measure the quantity of the nucleic acid(s) to be quantified. The invention also relates to a kit for carrying out said method.
摘要翻译：本发明涉及用于定量样品中一种或多种核酸的方法，例如：制备含有至少一种待定量核酸的样品，加入寡核苷酸探针，所述寡核苷酸探针包含可以与要定量的核酸特异性杂交或与要定量的核酸的共同序列杂交，在允许寡核苷酸探针与待定量的核酸杂交的条件下培养样品，将样品孵育允许杂交探针的扩展，每种情况下用作模板的核酸的步骤，从样品中除去非杂交探针，并定量杂交的寡核苷酸探针，以测量核酸的量量化。本发明还涉及一种用于执行所述方法的套件。

8. 发明授权

US08932831B2 Insertion of sequence elements into nucleic acids 有权
标题翻译：将序列元件插入核酸
公开(公告)号：US08932831B2
公开(公告)日：2015-01-13
申请号：US11744553
申请日：2007-05-04
申请人： Christian Korfhage , Holger Engel , Dirk Löffert , Ralf Peist
发明人： Christian Korfhage , Holger Engel , Dirk Löffert , Ralf Peist
IPC分类号： C07H21/00 , C12Q1/68 , C12P19/34 , C12N15/10
CPC分类号： C12P19/34 , C12N15/10 , C12Q1/6816 , C12Q2563/179 , C12Q2525/161 , C12Q2521/101
摘要： The present invention concerns a method for inserting one or more tag sequences into a nucleic acid characterized by the following steps: (a) preparation of a template nucleic acid; (b) hybridization of at least one anchor sequence of at least one anchor oligonucleotide with one sequence section of the template nucleic acid; and (c) synthesization of a new strand of nucleic acid, which is partially complementary to the template nucleic acid and which contains a sequence complementary to the non-hybridized portion of the anchor oligonucleotide, e.g. to at least one tag sequence, on its 3′ end.
摘要翻译：本发明涉及将一个或多个标签序列插入到以下步骤为特征的核酸的方法：（a）制备模板核酸; （b）将至少一种锚定寡核苷酸的至少一个锚定序列与模板核酸的一个序列部分杂交; 和（c）合成新的核酸链，其与模板核酸部分互补并且其含有与锚定寡核苷酸的非杂交部分互补的序列，例如，到其3'端的至少一个标签序列。

9. 发明申请

US20120107813A1 QUANTIFICATION OF NUCLEIC ACIDS 有权
标题翻译：核酸定量
公开(公告)号：US20120107813A1
公开(公告)日：2012-05-03
申请号：US13255696
申请日：2010-03-09
申请人： Nan Fang , Andreas Missel , Dirk Löffert
发明人： Nan Fang , Andreas Missel , Dirk Löffert
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6806 , C12Q2521/325
摘要： The invention relates to a method for the quantification of one or more nucleic acids in a sample. The method comprises the following steps: making a sample available which contains at least one nucleic acid to be quantified, adding an oligonucleotide probe to the sample, the oligonucleotide probe comprising a sequence which can specifically hybridize to the nucleic acid to be quantified or to a common sequence of the nucleic acids to be quantified, incubating the sample under conditions which allow the hybridization of the oligonucleotide probe to the nucleic acid(s) to be quantified, incubating the sample under conditions which allow the extension of hybridized probes, the nucleic acid(s) serving as a template in each case, removing the non-hybridized probes from the sample and quantifying the hybridized oligonucleotide probes to measure the quantity of the nucleic acid(s) to be quantified. The invention also relates to a kit for carrying out said method.
摘要翻译：本发明涉及用于定量样品中一种或多种核酸的方法。该方法包括以下步骤：制备含有至少一种待定量核酸的样品，向样品中加入寡核苷酸探针，所述寡核苷酸探针包含可与待定量的核酸特异性杂交的序列或寡核苷酸探针要量化的核酸的共同序列，在允许寡核苷酸探针杂交到待定量的核酸的条件下培养样品，在允许杂交探针的延伸的条件下培养样品，核酸在每种情况下用作模板，从样品中除去非杂交的探针并定量杂交的寡核苷酸探针以测量待定量的核酸的量。本发明还涉及一种用于执行所述方法的套件。

10. 发明授权

US08076069B2 Method for the treatment of a sample containing biomolecules 有权
标题翻译：用于处理含有生物分子的样品的方法
公开(公告)号：US08076069B2
公开(公告)日：2011-12-13
申请号：US12477813
申请日：2009-06-03
申请人： Christian Korfhage , Friederike Wilmer , Dirk Löffert , Ralf Himmelreich , Claudia Fritz , Kathleen Rieske
发明人： Christian Korfhage , Friederike Wilmer , Dirk Löffert , Ralf Himmelreich , Claudia Fritz , Kathleen Rieske
IPC分类号： C12Q1/68 , C07H21/00
CPC分类号： C12N1/06 , C12N15/1003 , C12Q1/6806 , G01N33/6803 , C12Q2531/113 , C12Q2527/137 , C12Q2527/125
摘要： The invention generally provides a method for the sample preparation for a subsequent preparation, processing or analysis method of a sample containing an at least one species of nucleic acid and/or one species of protein, whereby the method comprises the following steps: A) providing a sample which contains at least one species of a nucleic acid and/or of a protein, B) contacting the sample with a fluid or solid composition to produce a fluid sample preparation, whereby the composition contains at least a nitrogenous compound, which is selected from the group consisting of a) polyamines, b) amino acids, and oligo- and polypeptides, c) nitrogenous heterocyclic compounds, including homo- oder heteropolymers, which comprise these nitrogenous compounds, d) amines of the type R1R2NR3, whereby R1, R2 and R3 are chosen independently from one another from the group consisting of H, C1-C5-alkyl groups and aryl groups, whereby R1, R2 and R3 are not H simultaneously, e) carboxylic acid amides, f) inorganic ammonium salts, g) ammonium groups containing inner salt compounds, h) antibiotica binding nucleic acid, i) compounds which bind in the small cavity of the DNA, and mixtures of two or more of these compounds. The invention provides in particular a method for the lysis of a biological sample, and methods for the stabilization of biomolecules, a method for the decrease of inhibiting effects in a sample containing biomolecules and a differential masking method.
摘要翻译：本发明通常提供了用于样品制备用于随后的含有至少一种核酸和/或一种蛋白质的样品的制备，加工或分析方法的方法，其中所述方法包括以下步骤：A）提供包含至少一种核酸和/或蛋白质的样品，B）使样品与流体或固体组合物接触以产生流体样品制备物，其中所述组合物含有至少一种选自氮的化合物包括a）多胺，b）氨基酸和寡聚和多肽，c）包含这些含氮化合物的含氮杂环化合物，包括均相杂聚物，d）R1R2NR3型胺，其中R1，R2 和R 3彼此独立地选自H，C 1 -C 5烷基和芳基，其中R 1，R 2和R 3不同时为H，e）羧酸酰胺，f ）无机铵盐，g）含有内盐化合物的铵基，h）抗生素结合核酸，i）结合在DNA的小腔中的化合物，以及两种或更多种这些化合物的混合物。本发明特别提供了生物样品的裂解方法，以及用于稳定生物分子的方法，降低含有生物分子的样品中的抑制作用的方法和差示掩蔽法。

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