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1. 发明授权

US09217173B2 Method of polyadenylation and CDNA synthesis in a single reaction 有权
标题翻译：单次反应中聚腺苷酸化和CDNA合成的方法
公开(公告)号：US09217173B2
公开(公告)日：2015-12-22
申请号：US12377457
申请日：2007-08-13
申请人： Holger Engel , Subrahmanyam Yerramilli , Martin Kreutz , Dirk Loeffert , Christian Korfhage
发明人： Holger Engel , Subrahmanyam Yerramilli , Martin Kreutz , Dirk Loeffert , Christian Korfhage
IPC分类号： C12P19/34 , C12Q1/68
CPC分类号： C12Q1/6806 , C12Q1/6848 , C12Q2527/101 , C12Q2521/131 , C12Q2521/107
摘要： This invention relates to a process for synthesis of a cDNA in a sample, in an enzymatic reaction, wherein the process comprises the steps: simultaneous preparation 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; addition of a sample that comprises a ribonucleic acid; and incubation of the agents of the previous steps in one or more temperature steps, which are selected such that the first enzyme and the second enzyme show activity. The invention further relates to a reaction mixture that comprises 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, and optionally an anchor oligonucleotide. Moreover, the invention relates to a kit that comprises a corresponding reaction mixture.
摘要翻译：本发明涉及一种在酶促反应中合成样品中cDNA的方法，其中该方法包括以下步骤：同时制备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，至少缓冲液一个核糖核苷酸，至少一个脱氧核糖核苷酸，锚定寡核苷酸; 添加包含核糖核酸的样品; 以及先前步骤的试剂在一个或多个温度步骤中的温育，其被选择为使得第一酶和第二酶显示活性。本发明还涉及包含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，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和任选的锚定寡核苷酸的反应混合物。此外，本发明涉及包含相应反应混合物的试剂盒。

2. 发明申请

US20120202198A1 PROCESS FOR THE SYNTHESIS OF A CDNA IN A SAMPLE IN AN ENZYMATIC REACTION 有权
标题翻译：在一个酶反应中样品中合成CDNA的方法
公开(公告)号：US20120202198A1
公开(公告)日：2012-08-09
申请号：US12377457
申请日：2007-08-13
申请人： Holger Engel , Subrahmanyam Yerramilli , Martin Kreutz , Dirk Loeffert , Christian Korfhage
发明人： Holger Engel , Subrahmanyam Yerramilli , Martin Kreutz , Dirk Loeffert , Christian Korfhage
IPC分类号： C12P19/34 , C12N9/12 , C12Q1/68
CPC分类号： C12Q1/6806 , C12Q1/6848 , C12Q2527/101 , C12Q2521/131 , C12Q2521/107
摘要： This invention relates to a process for synthesis of a cDNA in a sample, in an enzymatic reaction, whereby the process comprises the steps: simultaneous preparation 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; addition of a sample that comprises a ribonucleic acid; and incubation of the agents of the previous steps in one or more temperature steps, which are selected such that the first enzyme and the second enzyme show activity. The invention further relates to a reaction mixture that comprises 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, and optionally an anchor oligonucleotide. Moreover, the invention relates to a kit that comprises a corresponding reaction mixture.
摘要翻译：本发明涉及在酶反应中合成样品中cDNA的方法，其中该方法包括以下步骤：同时制备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; 以及先前步骤的试剂在一个或多个温度步骤中的温育，其被选择为使得第一酶和第二酶显示活性。本发明还涉及包含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，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和任选的锚定寡核苷酸的反应混合物。此外，本发明涉及包含相应反应混合物的试剂盒。

3. 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。

4. 发明授权

US08486628B2 Method of normalized quantification of nucleic acids using anchor oligonucleotides and adapter oligonucleotides 失效
标题翻译：使用锚定寡核苷酸和衔接子寡核苷酸对核酸进行归一化定量的方法
公开(公告)号：US08486628B2
公开(公告)日：2013-07-16
申请号：US13388224
申请日：2010-07-30
申请人： Dirk Loeffert , Christian Korfhage , Holger Engel
发明人： Dirk Loeffert , Christian Korfhage , Holger Engel
IPC分类号： C12Q1/68 , C12P19/34 , C07H21/02 , C07H21/04
CPC分类号： C12Q1/6851 , C12Q2545/101 , C12Q2525/173 , C12Q2563/179 , C12Q2561/101
摘要： The present invention is related to normalized quantification of nucleic acids and to the normalization of quantities of nucleic acids in samples, e.g. mixtures of nucleic acids. The present invention relates to method for the normalization of the quantity of a nucleic acid to be quantified in a sample to the total quantity of nucleic acid in the sample; or to the total quantity of a specific class of nucleic acid in the sample.
摘要翻译：本发明涉及核酸的归一化定量和样品中核酸量的归一化，例如，核酸混合物。本发明涉及将样品中待定量的核酸的量归一化为样品中核酸总量的方法; 或相对于样品中特定类别的核酸的总量。

5. 发明申请

US20120190026A1 METHOD OF NORMALIZED QUANTIFICATION OF RNA 审中-公开
标题翻译： RNA的正规化定量方法
公开(公告)号：US20120190026A1
公开(公告)日：2012-07-26
申请号：US13387974
申请日：2010-07-30
申请人： Dirk Loeffert , Christian Korfhage , Holger Engel
发明人： Dirk Loeffert , Christian Korfhage , Holger Engel
IPC分类号： C12Q1/68 , G01N21/64
CPC分类号： C12Q1/6851 , C12Q2525/173 , C12Q2561/101 , C12Q2545/101 , C12Q2565/101 , C12Q2563/173 , C12Q2521/107
摘要： The present invention is related to normalized quantification of RNAs and to the normalization of quantities of RNAs in samples, e.g. mixtures of RNAs. The present invention relates to method for the normalization of the quantity of a RNA to be quantified in a sample to the total quantity of RNA in the sample; or to the total quantity of a specific class of RNA in the sample.
摘要翻译：本发明涉及RNA的归一化定量和样品中RNA的量的归一化。 RNA的混合物本发明涉及将样品中待定量的RNA的量归一化为样品中RNA总量的方法; 或样品中特定类别的RNA的总量。

6. 发明申请

US20120252014A1 METHOD OF NORMALIZED QUANTIFICATION OF NUCLEIC ACIDS USING ANCHOR OLIGONUCLEOTIDES AND ADAPTER OLIGONUCLEOTIDES 失效
标题翻译：使用锚定寡核苷酸和适配子寡核苷酸对核酸进行正规化定量的方法
公开(公告)号：US20120252014A1
公开(公告)日：2012-10-04
申请号：US13388224
申请日：2010-07-30
申请人： Dirk Loeffert , Christian Korfhage , Holger Engel
发明人： Dirk Loeffert , Christian Korfhage , Holger Engel
IPC分类号： G01N21/64
CPC分类号： C12Q1/6851 , C12Q2545/101 , C12Q2525/173 , C12Q2563/179 , C12Q2561/101
摘要： The present invention is related to normalized quantification of nucleic acids and to the normalization of quantities of nucleic acids in samples, e.g. mixtures of nucleic acids. The present invention relates to method for the normalization of the quantity of a nucleic acid to be quantified in a sample to the total quantity of nucleic acid in the sample; or to the total quantity of a specific class of nucleic acid in the sample.
摘要翻译：本发明涉及核酸的归一化定量和样品中核酸量的归一化，例如，核酸混合物。本发明涉及将样品中待定量的核酸的量归一化为样品中核酸总量的方法; 或相对于样品中特定类别的核酸的总量。

7. 发明申请

US20120190027A1 LIGATION-BASED METHOD OF NORMALIZED QUANTIFICATION OF NUCLEIC ACIDS 审中-公开
标题翻译：基于LIGATION的核酸正规化定量方法
公开(公告)号：US20120190027A1
公开(公告)日：2012-07-26
申请号：US13388228
申请日：2010-07-30
申请人： Dirk Loeffert , Christian Korfhage , Holger Engel
发明人： Dirk Loeffert , Christian Korfhage , Holger Engel
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6851 , C12Q2561/125 , C12Q2525/173
摘要： The present invention is related to normalized quantification of nucleic acids and to the normalization of quantities of nucleic acids in samples, e.g. mixtures of nucleic acids. The present invention relates to method for the normalization of the quantity of a nucleic acid to be quantified in a sample to the total quantity of nucleic acid in the sample; or to the total quantity of a specific class of nucleic acid in the sample.
摘要翻译：本发明涉及核酸的归一化定量和样品中核酸量的归一化，例如，核酸混合物。本发明涉及将样品中待定量的核酸的量归一化为样品中核酸总量的方法; 或相对于样品中特定类别的核酸的总量。

8. 发明授权

US10087482B2 Amplification of bisulfite-reacted nucleic acids 有权
公开(公告)号：US10087482B2
公开(公告)日：2018-10-02
申请号：US12865541
申请日：2009-02-06
申请人： Christian Korfhage , Dirk Loeffert , Ralf Peist , Nicolas Rudinger
发明人： Christian Korfhage , Dirk Loeffert , Ralf Peist , Nicolas Rudinger
IPC分类号： C12Q1/68 , C12Q1/6844
摘要： The present invention relates to a reaction mixture for the amplification of nucleic acids, the non-methylated cytosine bases of which have been converted to uracil bases by means of a bisulfition reaction. The invention also discloses methods for amplifying bisulfited nucleic acid and for determining the nucleic acid methylation state, and also kits based on the reaction mixture according to the invention.

9. 发明申请

US20110045542A1 AMPLIFICATION OF BISULFITE-REACTED NUCLEIC ACIDS 审中-公开
标题翻译：双相反应核酸的增殖
公开(公告)号：US20110045542A1
公开(公告)日：2011-02-24
申请号：US12865541
申请日：2009-02-06
申请人： Christian Korfhage , Dirk Loeffert , Ralf Peist , Nicolas Rudinger
发明人： Christian Korfhage , Dirk Loeffert , Ralf Peist , Nicolas Rudinger
IPC分类号： C12P19/34 , C12N9/10
CPC分类号： C12Q1/6844 , C12Q2523/125 , C12Q2527/137 , C12Q2531/119
摘要： The present invention relates to a reaction mixture for the amplification of nucleic acids, the non-methylated cytosine bases of which have been converted to uracil bases by means of a bisulfition reaction. The invention also discloses methods for amplifying bisulfited nucleic acid and for determining the nucleic acid methylation state, and also kits based on the reaction mixture according to the invention.
摘要翻译：本发明涉及用于扩增核酸的反应混合物，其非甲基化胞嘧啶碱基已经通过双硫酸反应转化成尿嘧啶碱基。本发明还公开了用于扩增亚硫酸核酸和确定核酸甲基化状态的方法，以及基于根据本发明的反应混合物的试剂盒。

10. 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。

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