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1. 发明申请

US20070264653A1 Method of identifying a biological sample for methylation analysis 审中-公开
标题翻译：识别甲基化分析生物样品的方法
公开(公告)号：US20070264653A1
公开(公告)日：2007-11-15
申请号：US11716207
申请日：2007-03-10
申请人： Kurt Berlin , Dimo Dietrich , Philipp Schatz , Michael Wandell , Antje Kluth , Reimo Tetzner
发明人： Kurt Berlin , Dimo Dietrich , Philipp Schatz , Michael Wandell , Antje Kluth , Reimo Tetzner
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6827 , C12Q1/6816 , C12Q1/6837 , C12Q2563/179 , C12Q2545/101 , C12Q2523/125
摘要： Aspects of the present invention relate to compositions and methods of identifying at least one biological sample in the field of methylation analysis. In particular aspects at least one biological sample is provided, at least one identifier is applied for each sample, the applied identifier(s) are detected or quantified, and the methylation analysis is performed. Additional aspects provide a methods for testing an experimental procedure. Additional aspects provide kits suitable for realizing the aspects of the invention.
摘要翻译：本发明的方面涉及在甲基化分析领域鉴定至少一种生物样品的组合物和方法。在特定方面，提供至少一个生物样品，每个样品应用至少一个标识符，检测或定量应用的标识符，并进行甲基化分析。其他方面提供了测试实验程序的方法。另外的方面提供适用于实现本发明方面的套件。

2. 发明申请

US20110059432A1 METHOD FOR PROVIDING DNA FRAGMENTS DERIVED FROM A REMOTE SAMPLE 审中-公开
标题翻译：用于提供从远程样品衍生的DNA片段的方法
公开(公告)号：US20110059432A1
公开(公告)日：2011-03-10
申请号：US11910887
申请日：2006-04-17
申请人： Matthias Ballhause , Kurt Berlin , Theo De Vos , Dimo Dietrich , Volker Liebenberg , Catherine Lofton-Day , Joe Lograsso , Jennifer Maas , Fabian Model , Matthias Schuster , Andrew Z. Sledziewski , Reimo Tetzner
发明人： Matthias Ballhause , Kurt Berlin , Theo De Vos , Dimo Dietrich , Volker Liebenberg , Catherine Lofton-Day , Joe Lograsso , Jennifer Maas , Fabian Model , Matthias Schuster , Andrew Z. Sledziewski , Reimo Tetzner
IPC分类号： C12Q1/68 , C07H21/04 , C12P19/34 , G01N33/48
摘要： Aspects of the present invention relate to compositions and methods for providing DNA fragments from a remote sample. In particular aspects a remote sample comprising DNA is provided, DNA is isolated from the remote sample, and the isolated DNA is treated in a way which allows differentiation of methylated and unmethylated cytosine. Additional, particular embodiments provide compositions and methods for methylation analysis of DNA derived from a remote sample. Other aspects provide for compositions and methods of whole genome amplification of bisulfite treated DNA.
摘要翻译：本发明的方面涉及从远程样品提供DNA片段的组合物和方法。在特定方面，提供了包含DNA的远程样品，从远程样品中分离DNA，并以允许甲基化和非甲基化胞嘧啶分化的方式处理分离的DNA。另外，具体实施方案提供了用于从远程样品衍生的DNA的甲基化分析的组合物和方法。其他方面提供了亚硫酸氢盐处理的DNA的全基因组扩增的组合物和方法。

3. 发明授权

US10731215B2 Method for determining the presence or absence of methylation in a sample 有权
公开(公告)号：US10731215B2
公开(公告)日：2020-08-04
申请号：US11910887
申请日：2006-04-17
申请人： Matthias Ballhause , Kurt Berlin , Theo De Vos , Dimo Dietrich , Volker Liebenberg , Catherine Lofton-Day , Joe Lograsso , Jennifer Maas , Fabian Model , Matthias Schuster , Andrew Z. Sledziewski , Reimo Tetzner
发明人： Matthias Ballhause , Kurt Berlin , Theo De Vos , Dimo Dietrich , Volker Liebenberg , Catherine Lofton-Day , Joe Lograsso , Jennifer Maas , Fabian Model , Matthias Schuster , Andrew Z. Sledziewski , Reimo Tetzner
IPC分类号： C12Q1/68 , C12Q1/6883 , C12N15/10 , C12Q1/6806 , C07H21/04
摘要： Aspects of the present invention relate to compositions and methods for providing DNA fragments from a remote sample. In particular aspects a remote sample comprising DNA is provided, DNA is isolated from the remote sample, and the isolated DNA is treated in a way which allows differentiation of methylated and unmethylated cytosine. Additional, particular embodiments provide compositions and methods for methylation analysis of DNA derived from a remote sample. Other aspects provide for compositions and methods of whole genome amplification of bisulfite treated DNA. Other aspects provide methods for determining the presence or absence of methylation of at least one cytosine, or a series of cytosines in cis, in human DNA of a blood sample, a plasma sample, a serum sample or a urine sample from a human individual.

4. 发明申请

US20090075251A1 Method for analysis of cytosine methylation 审中-公开
标题翻译：胞嘧啶甲基化分析方法
公开(公告)号：US20090075251A1
公开(公告)日：2009-03-19
申请号：US10594013
申请日：2005-03-24
申请人： Dimo Dietrich , Philipp Schatz , Matthias Schuster , Antje Kluth
发明人： Dimo Dietrich , Philipp Schatz , Matthias Schuster , Antje Kluth
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6858 , C12Q2525/143 , C12Q2523/125
摘要： A method for the analysis of cytosine methylations in DNA is described. Here, the DNA to be investigated is first chemically or enzymatically converted. Then a promoter is introduced into the DNA. After this, the DNA is converted to RNA. The methylation pattern of the DNA can be concluded in different ways by means of an analysis of the RNA. The RNA is preferably fragmented chemically or enzymatically prior to the analysis, whereby the fragmenting results depend on the methylation pattern of the DNA. The method according to the invention is particularly suitable for the diagnosis and prognosis of cancer disorders and other diseases associated with a modification of the methylation pattern.
摘要翻译：描述了分析DNA中胞嘧啶甲基化的方法。这里，待研究的DNA首先被化学或酶转化。然后将启动子引入DNA。之后，将DNA转化为RNA。 DNA的甲基化模式可以通过RNA的分析以不同的方式得出结论。优选在分析之前将RNA片段化学或酶促，由此片段化结果取决于DNA的甲基化模式。根据本发明的方法特别适合于与甲基化模式的改变相关的癌症病症和其它疾病的诊断和预后。

5. 发明授权

US08679745B2 Method for providing DNA fragments derived from an archived sample 有权
标题翻译：提供衍生自归档样品的DNA片段的方法
公开(公告)号：US08679745B2
公开(公告)日：2014-03-25
申请号：US11664367
申请日：2005-09-30
申请人： Matthias Ballhause , Kurt Berlin , Dimo Dietrich , Antje Kluth Lukas , Matthias Schuster , Ute Wagner , Reinhold Wasserkort , Heike Ziebarth
发明人： Matthias Ballhause , Kurt Berlin , Dimo Dietrich , Antje Kluth Lukas , Matthias Schuster , Ute Wagner , Reinhold Wasserkort , Heike Ziebarth
IPC分类号： C12Q1/68 , C12P19/34 , B01L3/00
CPC分类号： C12Q1/6806 , C12N15/1003 , C12Q1/6858 , C12Q1/686 , C12Q2523/125 , C12Q2527/137 , C12Q2527/149 , C12Q2531/107
摘要： Aspects of the present invention relate to compositions and methods for providing DNA fragments from an archived sample (e.g., paraffin-embedded and/or fixed-tissue biopsies, etc). Particular aspects provide methods whereby high yields of DNA are isolated as well as a substantial portion of the DNA consists of long DNA fragments, and where the isolated genomic DNA is free of associated or cross-linked contaminants like proteins, peptides, amino acids or RNA. The methods are facile, cost-effective, and are characterized by high reproducibility and reliability. Particular aspects provide methods for providing DNA fragments derived from an archived sample, wherein the yield of DNA before, for example, an amplification step is at least 20%, and amplicons up to a length of about 1,000 base pairs are amplifiable.
摘要翻译：本发明的方面涉及从归档样品（例如石蜡包埋和/或固定组织活组织检查等）提供DNA片段的组合物和方法。具体方面提供了分离DNA的高产率以及大部分DNA由长DNA片段组成并且分离的基因组DNA不含相关或交联的污染物如蛋白质，肽，氨基酸或RNA的方法。该方法简便，成本效益高，重现性和可靠性高。特定方面提供用于提供衍生自归档样品的DNA片段的方法，其中在例如扩增步骤之前的DNA的产量为至少20％，并且可扩增至约1000个碱基对的扩增子。

6. 发明授权

US07700282B2 Method for the carry-over protection in DNA amplification systems targeting methylation analysis achieved by a modified pre-treatment of nucleic acids 有权
标题翻译：用于DNA扩增系统中的遗传保护的方法，其靶向通过核酸的修饰预处理实现的甲基化分析
公开(公告)号：US07700282B2
公开(公告)日：2010-04-20
申请号：US11248721
申请日：2005-10-11
申请人： Reimo Tetzner , Dimo Dietrich
发明人： Reimo Tetzner , Dimo Dietrich
IPC分类号： C12Q1/70 , C12P19/34
CPC分类号： C12Q1/6876 , C12P19/34 , C12Q1/6806 , C12Q1/6827 , C12Q1/6848 , C12Q1/6883 , C12Q1/6886 , C12Q2600/156 , C12Q2523/125 , C12Q2521/531 , C12Q2531/113
摘要： Particular aspects provide methods for specific amplification of template DNA in the presence of potentially contaminating PCR products from previous amplification experiments. Particular embodiments comprise, in a first step, contacting DNA with a bisulfite solution, which sulfonates unmethylated (but not methylated) cytosines, resulting in cytosine deamination and generation of sulfonated uracil. Such sulfonation protects the template nucleic acid from being a target for the enzyme uracil-DNA-glycosylase (UNG), whereas any contaminating DNA, which contains unprotected unsulfonated or desulfonated uracils, is degraded enzymatically while the UNG is active. After UNG treatment and inactivation thereof, the sulfonated uracil bases are converted into uracil by desulfonation. Such aspects have substantial utility for decontamination of nucleic acid samples; e.g., for avoiding amplification of ‘carry over products’ in the context of DNA methylation analysis. In further aspects, the inventive methods can be generally used as simplified methods of bisulfite treatment.
摘要翻译：在以前的扩增实验中存在可能污染的PCR产物的情况下，特定方面提供用于特异性扩增模板DNA的方法。具体实施方案包括在第一步中使DNA与亚硫酸氢盐溶液接触，所述亚硫酸氢盐溶液磺化非甲基化（但不是甲基化的）胞嘧啶，导致胞嘧啶脱氨基并产生磺化的尿嘧啶。这种磺化保护模板核酸不是酶尿嘧啶-DNA-糖基化酶（UNG）的靶标，而任何含有未保护的未磺化或脱磺酸尿嘧啶的污染性DNA在UNG活性时被酶促降解。在UNG处理和灭活后，通过脱磺酸将磺化的尿嘧啶碱基转化成尿嘧啶。这些方面对于核酸样品的净化具有实质性的实用性; 例如，为了避免在DNA甲基化分析的上下文中“携带产物”的扩增。在另外的方面，本发明的方法通常可以用作亚硫酸氢盐处理的简化方法。

7. 发明授权

US08753810B2 Method for the carry-over protection in DNA amplification systems targeting methylation analysis achieved by a modified pre-treatment of nucleic acids 有权
标题翻译：用于DNA扩增系统中的遗传保护的方法，其靶向通过核酸的修饰预处理实现的甲基化分析
公开(公告)号：US08753810B2
公开(公告)日：2014-06-17
申请号：US12709300
申请日：2010-02-19
申请人： Reimo Tetzner , Dimo Dietrich
发明人： Reimo Tetzner , Dimo Dietrich
IPC分类号： C12Q1/68 , C12P19/34
CPC分类号： C12Q1/6876 , C12P19/34 , C12Q1/6806 , C12Q1/6827 , C12Q1/6848 , C12Q1/6883 , C12Q1/6886 , C12Q2600/156 , C12Q2523/125 , C12Q2521/531 , C12Q2531/113
摘要： Particular aspects provide methods for specific amplification of template DNA in the presence of potentially contaminating PCR products from previous amplification experiments. Particular embodiments comprise, in a first step, contacting DNA with a bisulfite solution, which sulfonates unmethylated (but not methylated) cytosines, resulting in cytosine deamination and generation of sulfonated uracil. Such sulfonation protects the template nucleic acid from being a target for the enzyme uracil-DNA-glycosylase (UNG), whereas any contaminating DNA, which contains unprotected unsulfonated or desulfonated uracils, is degraded enzymatically while the UNG is active. After UNG treatment and inactivation thereof, the sulfonated uracil bases are converted into uracil by desulfonation. Such aspects have substantial utility for decontamination of nucleic acid samples; e.g., for avoiding amplification of ‘carry over products’ in the context of DNA methylation analysis. In further aspects, the inventive methods can be generally used as simplified methods of bisulfite treatment.
摘要翻译：在以前的扩增实验中存在可能污染的PCR产物的情况下，特定方面提供用于特异性扩增模板DNA的方法。具体实施方案包括在第一步中使DNA与亚硫酸氢盐溶液接触，所述亚硫酸氢盐溶液磺化非甲基化（但不是甲基化的）胞嘧啶，导致胞嘧啶脱氨基并产生磺化的尿嘧啶。这种磺化保护模板核酸不是酶尿嘧啶-DNA-糖基化酶（UNG）的靶标，而任何含有未保护的未磺化或脱磺酸尿嘧啶的污染性DNA在UNG活性时被酶促降解。在UNG处理和灭活后，通过脱磺酸将磺化的尿嘧啶碱基转化成尿嘧啶。这些方面对于核酸样品的净化具有实质性的实用性; 例如，为了避免在DNA甲基化分析的上下文中“携带产物”的扩增。在另外的方面，本发明的方法通常可以用作亚硫酸氢盐处理的简化方法。

8. 发明申请

US20110027834A1 Carry-Over Protection in Enzyme-Based Dna Amplification Systems Targeting Methylation Analysis 审中-公开
标题翻译：基于甲基化分析的酶基Dna扩增系统中的保护
公开(公告)号：US20110027834A1
公开(公告)日：2011-02-03
申请号：US12308149
申请日：2007-06-08
申请人： Reimo Tetzner , Dimo Dietrich
发明人： Reimo Tetzner , Dimo Dietrich
IPC分类号： C12P19/34 , C12N9/00 , C07H21/04
CPC分类号： C12Q1/6848 , C12Q2525/131 , C12Q2523/125
摘要： The invention refers to a method for providing a decontaminated template nucleic acid for enzymatic amplification reactions suitable for DNA methylation analysis. This method is characterized by the following steps: a) incubating a nucleic acid with a chemical reagent or an enzyme-containing solution, whereby the unmethylated cytosine bases are converted into uracil bases, b) mixing the template nucleic acid from step a) with the components required for an enzyme-mediated amplification reaction, including at least two oligonucleotides, whereby at least one of said oligonucleotides comprises i) at least one sequence part that hybridizes with a sequence of the template nucleic acid to be amplified, and ii) at least one sequence part that constitutes a recognition site for a DNA cleaving enzyme that cleaves DNA downstream of said recognition site and c) adding to this mixture a DNA cleaving enzyme, which specifically binds to the at least one sequence part that is a recognition site, and d) incubating the mixture, whereby nucleic acids containing said recognition site for a DNA cleaving enzyme are degraded.
摘要翻译：本发明涉及一种用于提供适用于DNA甲基化分析的酶扩增反应的去污染模板核酸的方法。该方法的特征在于以下步骤：a）将核酸与化学试剂或含酶溶液一起孵育，由此将未甲基化的胞嘧啶碱基转化成尿嘧啶碱基，b）将来自步骤a）的模板核酸与酶介导的扩增反应所需的组分，包括至少两个寡核苷酸，其中至少一个所述寡核苷酸包含i）至少一个与待扩增的模板核酸序列杂交的序列部分，和ii）至少构成用于切割所述识别位点下游的DNA的DNA切割酶的识别位点的一个序列部分，和c）向该混合物中加入特异性结合作为识别位点的至少一个序列部分的DNA切割酶，以及 d）孵育混合物，由此含有用于DNA切割酶的所述识别位点的核酸被降解。

9. 发明申请

US20110003292A1 METHODS AND NUCLEIC ACIDS FOR ANALYSES OF CELL PROLIFERATIVE DISORDERS 审中-公开
标题翻译：方法和细胞增殖性疾病分析的核酸
公开(公告)号：US20110003292A1
公开(公告)日：2011-01-06
申请号：US12747584
申请日：2008-12-11
申请人： Dimo Dietrich , Volker Liebenberg , Reimo Tetzner , Juergen Distler , Joern Lewin , Thomas Schlegel
发明人： Dimo Dietrich , Volker Liebenberg , Reimo Tetzner , Juergen Distler , Joern Lewin , Thomas Schlegel
IPC分类号： C12Q1/68 , G01N33/53 , C12M1/34
CPC分类号： C12Q1/6886 , C12Q2600/154 , C12Q2600/156 , C12Q2600/158 , C12Q2600/16
摘要： The invention provides methods, nucleic acids and kits for detecting lung carcinoma. The invention discloses genomic (FOXL2, ONECUT1, TFAP2E, EN2-2, EN2-3, SHOX2-2 and BARHL2) sequences the methylation patterns of which have utility for the improved detection of said disorder, thereby enabling the improved diagnosis and treatment of patients.
摘要翻译：本发明提供用于检测肺癌的方法，核酸和试剂盒。本发明公开了基因组（FOXL2，ONECUT1，TFAP2E，EN2-2，EN2-3，SHOX2-2和BARHL2）序列，其甲基化模式可用于改善所述病症的检测，从而能够改善患者的诊断和治疗。

10. 发明申请

US20100216152A1 Method for the Carry-Over Protection in DNA Amplification Systems Targeting Methylation Analysis Achieved by a Modified Pre-Treatment of Nucleic Acids 有权
标题翻译：靶向通过核酸修饰预处理实现的甲基化分析的DNA扩增系统中的转移保护方法
公开(公告)号：US20100216152A1
公开(公告)日：2010-08-26
申请号：US12709300
申请日：2010-02-19
申请人： Reimo Tetzner , Dimo Dietrich
发明人： Reimo Tetzner , Dimo Dietrich
IPC分类号： C12Q1/68 , C12P19/34 , C12N9/00
CPC分类号： C12Q1/6876 , C12P19/34 , C12Q1/6806 , C12Q1/6827 , C12Q1/6848 , C12Q1/6883 , C12Q1/6886 , C12Q2600/156 , C12Q2523/125 , C12Q2521/531 , C12Q2531/113
摘要： Particular aspects provide methods for specific amplification of template DNA in the presence of potentially contaminating PCR products from previous amplification experiments. Particular embodiments comprise, in a first step, contacting DNA with a bisulfite solution, which sulfonates unmethylated (but not methylated) cytosines, resulting in cytosine deamination and generation of sulfonated uracil. Such sulfonation protects the template nucleic acid from being a target for the enzyme uracil-DNA-glycosylase (UNG), whereas any contaminating DNA, which contains unprotected unsulfonated or desulfonated uracils, is degraded enzymatically while the UNG is active. After UNG treatment and inactivation thereof, the sulfonated uracil bases are converted into uracil by desulfonation. Such aspects have substantial utility for decontamination of nucleic acid samples; e.g., for avoiding amplification of ‘carry over products’ in the context of DNA methylation analysis. In further aspects, the inventive methods can be generally used as simplified methods of bisulfite treatment.
摘要翻译：在以前的扩增实验中存在可能污染的PCR产物的情况下，特定方面提供用于特异性扩增模板DNA的方法。具体实施方案包括在第一步中使DNA与亚硫酸氢盐溶液接触，所述亚硫酸氢盐溶液磺化非甲基化（但不是甲基化的）胞嘧啶，导致胞嘧啶脱氨基并产生磺化的尿嘧啶。这种磺化保护模板核酸不是酶尿嘧啶-DNA-糖基化酶（UNG）的靶标，而含有未保护的未磺化或脱磺酸尿嘧啶的任何污染性DNA在UNG活性时被酶促降解。在UNG处理和灭活后，通过脱磺酸将磺化的尿嘧啶碱基转化成尿嘧啶。这些方面对于核酸样品的净化具有实质性的实用性; 例如，为了避免在DNA甲基化分析的上下文中“携带产物”的扩增。在另外的方面，本发明的方法通常可以用作亚硫酸氢盐处理的简化方法。

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