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11. 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。

12. 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。

13. 发明申请

US20080096199A1 Method for the Carry-Over Protection In Dna Amplification Systems Targeting Methylation Analysis Achieved by a Modified Pre-Treatment of Nucleic Acids 审中-公开
标题翻译： Dna扩增系统中转移保护的方法靶向通过核酸修饰预处理实现的甲基化分析
公开(公告)号：US20080096199A1
公开(公告)日：2008-04-24
申请号：US11665196
申请日：2005-10-11
申请人： Reimo Tetzner , Dimo Dietrich
发明人： Reimo Tetzner , Dimo Dietrich
IPC分类号： C12Q1/68 , 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： Disclosed is a method for the specific amplification of template DNA in the presence of potentially contaminating PCR products from previous amplification experiments. In the first step DNA is contacted with a bisulfite solution, which reacts with unmethylated cytosines but not with methylated cytosines, by sulfonating them. This results in deamination of the cytosine whereby sulfonated uracil is generated. Such sulfonation protects the template nucleic acid from being a target for the enzyme UNG. Any contaminating DNA, which contains unprotected unsulfonated or desulfonated uracils is degraded enzymatically while UNG is active. After UNG treatment and inactivation, the sulfonated uracil bases are converted into uracil by desulfonation. This method is useful for decontamination of nucleic acid samples, or rather for avoiding amplification of ‘carry over products’ in particular in the context of DNA methylation analysis. Furthermore it can be used as a simplified method of bisulfite treatment in general.
摘要翻译：公开了在先前扩增实验中存在可能污染的PCR产物的情况下特异性扩增模板DNA的方法。在第一步中，DNA与亚硫酸氢盐溶液接触，其与未甲基化的胞嘧啶反应而不与甲基化的胞嘧啶反应，通过磺化它们。这导致胞嘧啶脱氨，从而产生磺化的尿嘧啶。这种磺化保护模板核酸不是酶UNG的靶标。含有未受保护的未磺化或脱磺酸尿嘧啶的任何污染性DNA在酶活性下降，而UNG活性。在UNG处理和灭活后，磺化的尿嘧啶碱基通过脱磺酸转化成尿嘧啶。该方法可用于核酸样品的去污，或者用于避免在DNA甲基化分析的背景下特别是“携带产物”的扩增。此外，它可以用作一般的亚硫酸氢盐处理的简化方法。

14. 发明申请

US20060115835A1 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：US20060115835A1
公开(公告)日：2006-06-01
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： C12Q1/68 , C07H21/04 , 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。

15. 发明授权

US08771939B2 Method for methylation analysis of nucleic acid 有权
标题翻译：核酸甲基化分析方法
公开(公告)号：US08771939B2
公开(公告)日：2014-07-08
申请号：US12310051
申请日：2007-08-01
申请人： Reimo Tetzner , Joern Lewin
发明人： Reimo Tetzner , Joern Lewin
IPC分类号： C12Q1/68 , C12P19/34 , C07H21/02 , C07H21/04
CPC分类号： C12Q1/6858 , C12Q2537/113 , C12Q2535/119 , C12Q2523/125
摘要： The present invention relates to a method for methylation analysis. It comprises the providing of a double stranded nucleic acid; its conversion, whereby unmethylated bases become distinguishable in their base-pairing behavior from methylated bases, and the analysis of both of the converted nucleic acid strands.
摘要翻译：本发明涉及甲基化分析方法。它包括提供双链核酸; 其转化，其中未甲基化的碱基在其碱基配对行为与甲基化碱基之间变得可区分，以及两个转化的核酸链的分析。

16. 发明授权

US09695478B2 Methods and nucleic acids for the analyses of cellular proliferative disorders 有权
公开(公告)号：US09695478B2
公开(公告)日：2017-07-04
申请号：US12779856
申请日：2010-05-13
申请人： Catherine E. Lofton-Day , Andrew Z. Sledziewski , Ralf Lesche , Matthias Schuster , Juergen Distler , Reimo Tetzner , Thomas Hildmann , Fabian Model , Xiaoling Song
发明人： Catherine E. Lofton-Day , Andrew Z. Sledziewski , Ralf Lesche , Matthias Schuster , Juergen Distler , Reimo Tetzner , Thomas Hildmann , Fabian Model , Xiaoling Song
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6886 , C12Q1/6806 , C12Q2600/106 , C12Q2600/112 , C12Q2600/154 , C12Q2600/158
摘要： The invention provides methods, nucleic acids and kits for detecting, or for detecting and distinguishing between or among liver cell proliferative disorders or for detecting, or for detecting and distinguishing between or among colorectal cell proliferative disorders. The invention discloses genomic sequences the methylation patterns of which have utility for the improved detection of and differentiation between said class of disorders, thereby enabling the improved diagnosis and treatment of patients.

17. 发明申请

US20080311627A1 Compositions and Methods for Preventing Carry-Over Contamination in Nucleic Acid Amplification Reactions 审中-公开
标题翻译：防止核酸扩增反应中携带污染的组合物和方法
公开(公告)号：US20080311627A1
公开(公告)日：2008-12-18
申请号：US11663420
申请日：2005-06-17
申请人： Reimo Tetzner , Kurt Berlin , Juergen Distler
发明人： Reimo Tetzner , Kurt Berlin , Juergen Distler
IPC分类号： C12P19/34
CPC分类号： C12Q1/6848 , C12Q2527/101 , C12Q2521/301
摘要： Particular aspects provide methods for the specific amplification of template DNA in the presence of potentially contaminating PCR products from previous amplification experiments, but wherein the contaminating prior reaction amplificates (carry-over contaminants) are rendered non-amplifiable, based on use of particular contaminant degradation enzymes, and use of at least one primer that is fully complementary to any contaminating prior reaction amplificate nucleic acid, but contains a mismatch with the correspond sequence of the sample template nucleic acid. Additional aspects provide a method for the specific amplification of single-stranded sample template DNA in the presence of potentially double-stranded carry-over products. Further aspects provide methods comprising use of a template-dependent thermostable DNA polymerase enzyme suitable for incorporating ribonucleotides as well as deoxy-nucleotides to provide a chimeric amplificate. After digestion with an RNase, any contaminating prior chimeric amplificate, the RNase is inactivated. These aspects are surprisingly effective alternatives to the carry over protection system known as UNG system, and other art-recognized methods.
摘要翻译：特定方面提供了在来自先前扩增实验的潜在污染性PCR产物存在下对模板DNA进行特异性扩增的方法，但其中基于使用特定污染物降解使污染的先前反应扩增（携带污染物）变得不可扩增酶，以及使用与任何污染的先前反应扩增核酸完全互补的至少一个引物，但是含有与样品模板核酸的相应序列错配。另外的方面提供了在存在潜在双链携带产物的情况下单链样品模板DNA的特异性扩增的方法。另外的方面提供了包括使用适合于掺入核糖核苷酸的模板依赖性热稳定性DNA聚合酶以及脱氧核苷酸以提供嵌合扩增的方法。用RNase消化后，任何污染的先前的嵌合扩增，核糖核酸酶被灭活。这些方面对于被称为UNG系统的承载保护系统以及其他本领域公认的方法是惊人的有效的替代方案。

18. 发明申请

US20060286576A1 Methods and nucleic acids for the analyses of cellular proliferative disorders 有权
标题翻译：用于分析细胞增殖性疾病的方法和核酸
公开(公告)号：US20060286576A1
公开(公告)日：2006-12-21
申请号：US11405322
申请日：2006-04-17
申请人： Cathy Lofton-Day , Andrew Sledziewski , Ralf Lesche , Matthias Schuster , Juergen Distler , Reimo Tetzner , Thomas Hildmann , Fabian Model , Xiaoling Song
发明人： Cathy Lofton-Day , Andrew Sledziewski , Ralf Lesche , Matthias Schuster , Juergen Distler , Reimo Tetzner , Thomas Hildmann , Fabian Model , Xiaoling Song
IPC分类号： C12Q1/68 , C07H21/04 , C12P19/34
CPC分类号： C12Q1/6886 , C12Q1/6806 , C12Q2600/106 , C12Q2600/112 , C12Q2600/154 , C12Q2600/158
摘要： The invention provides methods, nucleic acids and kits for detecting, or for detecting and distinguishing between or among liver cell proliferative disorders or for detecting, or for detecting and distinguishing between or among colorectal cell proliferative disorders. The invention discloses genomic sequences the methylation patterns of which have utility for the improved detection of and differentiation between said class of disorders, thereby enabling the improved diagnosis and treatment of patients.
摘要翻译：本发明提供用于检测或检测和鉴别肝细胞增殖性疾病或用于检测或检测和鉴别结直肠细胞增殖性疾病之间或之间的方法，核酸和试剂盒。本发明公开了基因组序列，其甲基化模式可用于改善所述疾病类别的检测和分化，从而能够改善患者的诊断和治疗。

19. 发明授权

US08623599B2 Method for methylation analysis 有权
标题翻译：甲基化分析方法
公开(公告)号：US08623599B2
公开(公告)日：2014-01-07
申请号：US12451943
申请日：2008-06-06
申请人： Theo Devos , Cathy Lofton-Day , Andrew Sledziewski , Fabian Model , Michael Krouse , Jesse Ho , Matthias Schuster , Juergen Distler , Reimo Tetzner
发明人： Theo Devos , Cathy Lofton-Day , Andrew Sledziewski , Fabian Model , Michael Krouse , Jesse Ho , Matthias Schuster , Juergen Distler , Reimo Tetzner
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6813 , C12Q1/6806 , C12Q1/6827 , C12Q1/6844 , C12Q2523/125
摘要： Aspects of the invention relate to composition and methods for the providing of DNA for methylation analysis that is in particular suitable to be applied in reference laboratories. Further 5 aspects of the invention relate to composition and methods for the highly specific and sensitive methylation analysis of the Septin 9 gene also in particular suitable to be applied in reference laboratories.
摘要翻译：本发明的方面涉及提供用于甲基化分析的DNA的组合物和方法，其特别适用于参考实验室。本发明的另外5个方面涉及用于Septin 9基因的高度特异性和敏感的甲基化分析的组合物和方法，特别适用于参考实验室。

20. 发明授权

US07749702B2 Methods and nucleic acids for the analyses of cellular proliferative disorders 有权
标题翻译：用于分析细胞增殖性疾病的方法和核酸
公开(公告)号：US07749702B2
公开(公告)日：2010-07-06
申请号：US11405322
申请日：2006-04-17
申请人： Catherine E. Lofton-Day , Andrew Z. Sledziewski , Ralf Lesche , Matthias Schuster , Juergen Distler , Reimo Tetzner , Thomas Hildmann , Fabian Model , Xiaoling Song
发明人： Catherine E. Lofton-Day , Andrew Z. Sledziewski , Ralf Lesche , Matthias Schuster , Juergen Distler , Reimo Tetzner , Thomas Hildmann , Fabian Model , Xiaoling Song
IPC分类号： C12Q1/68 , C12Q1/44 , C12P19/34 , C07H21/04
CPC分类号： C12Q1/6886 , C12Q1/6806 , C12Q2600/106 , C12Q2600/112 , C12Q2600/154 , C12Q2600/158
摘要： The invention provides methods, nucleic acids and kits for detecting, or for detecting and distinguishing between or among liver cell proliferative disorders or for detecting, or for detecting and distinguishing between or among colorectal cell proliferative disorders. The invention discloses genomic sequences the methylation patterns of which have utility for the improved detection of and differentiation between said class of disorders, thereby enabling the improved diagnosis and treatment of patients.
摘要翻译：本发明提供用于检测或检测和鉴别肝细胞增殖性疾病或用于检测或检测和鉴别结直肠细胞增殖性疾病之间或之间的方法，核酸和试剂盒。本发明公开了基因组序列，其甲基化模式可用于改善所述疾病类别的检测和分化，从而能够改善患者的诊断和治疗。

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