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1. 发明申请

US20110250601A1 Bisulfite Conversion of DNA 有权
标题翻译：亚硫酸氢盐转化DNA
公开(公告)号：US20110250601A1
公开(公告)日：2011-10-13
申请号：US13159323
申请日：2011-06-13
申请人： Kurt Berlin , Matthias Ballhause , Karen Cardon
发明人： Kurt Berlin , Matthias Ballhause , Karen Cardon
IPC分类号： C12Q1/68 , C07H21/04 , G01N33/50
CPC分类号： C12Q1/6827 , C12Q2533/101 , Y10T436/143333 , C12Q2527/125 , C12Q2523/125 , C12Q2531/125
摘要： The present invention provides an improved method for the bisulfite conversion of DNA, and facilitates the analysis of cytosine methylation of genomic DNA. Novel combinations of denaturing solvents, new reaction conditions and new purification methods provide surprisingly efficacious methods for bisulfite conversion of DNA relative to prior art methods. The converted DNA may subsequently be analyzed by many different methods.
摘要翻译：本发明提供了DNA的亚硫酸氢盐转化的改进方法，并且有助于分析基因组DNA的胞嘧啶甲基化。相对于现有技术的方法，变性溶剂，新反应条件和新的纯化方法的新型组合为DNA的亚硫酸氢盐转化提供了惊人的有效方法。随后可以通过许多不同的方法分析转化的DNA。

2. 发明申请

US20110159491A1 Method for detecting cytosine methylation in DNA samples 审中-公开
标题翻译： DNA样品中胞嘧啶甲基化检测方法
公开(公告)号：US20110159491A1
公开(公告)日：2011-06-30
申请号：US12925843
申请日：2010-10-29
申请人： Alexander Olek , Kurt Berlin
发明人： Alexander Olek , Kurt Berlin
IPC分类号： C12Q1/68
CPC分类号： C12Q1/686 , C12Q1/6816 , C12Q1/6827 , C12Q1/6837 , C12Q1/6872 , C12Q2523/125 , C12Q2565/519 , C12Q2565/627
摘要： Described is a method for detecting 5-methylcytosine in genomic DNA samples. First, a genomic DNA from a DNA sample is chemically converted with a reagent, 5-methylcytosine and cytosine reacting differently, and the pretreated DNA is subsequently amplified using a polymerase and at least one primer. In the next step, the amplified genomic DNA is hybridized to at least one oligonucleotide, forming a duplex, and said oligonucleotide is elongated by at least one nucleotide, the nucleotide carrying a detectable label, and the elongation depending on the methylation status of the specific cytosine in the genomic DNA sample. In the next step, the elongated oligonucleotides are analyzed for the presence of the label.
摘要翻译：描述了在基因组DNA样品中检测5-甲基胞嘧啶的方法。首先，DNA样品的基因组DNA用试剂化学转化，5-甲基胞嘧啶和胞嘧啶的反应不同，然后使用聚合酶和至少一种引物扩增预处理的DNA。在下一步骤中，将扩增的基因组DNA与至少一种形成双链体的寡核苷酸杂交，并且所述寡核苷酸由至少一个核苷酸延伸，所述核苷酸携带可检测标记，延伸取决于具体的甲基化状态基因组DNA样品中的胞嘧啶。在下一步骤中，分析细长寡核苷酸的标记物的存在。

3. 发明授权

US07968295B2 Bisulfite conversion of DNA 有权
标题翻译：亚硫酸氢盐转化DNA
公开(公告)号：US07968295B2
公开(公告)日：2011-06-28
申请号：US12410389
申请日：2009-03-24
申请人： Kurt Berlin , Matthias Ballhause , Karen Cardon
发明人： Kurt Berlin , Matthias Ballhause , Karen Cardon
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6827 , C12Q2533/101 , Y10T436/143333 , C12Q2527/125 , C12Q2523/125 , C12Q2531/125
摘要： The present invention provides an improved method for the bisulfite conversion of DNA, and facilitates the analysis of cytosine methylation of genomic DNA. Novel combinations of denaturing solvents, new reaction conditions and new purification methods provide surprisingly efficacious methods for bisulfite conversion of DNA relative to prior art methods. The converted DNA may subsequently be analyzed by many different methods.
摘要翻译：本发明提供了DNA的亚硫酸氢盐转化的改进方法，并且有助于分析基因组DNA的胞嘧啶甲基化。相对于现有技术的方法，变性溶剂，新反应条件和新的纯化方法的新型组合为DNA的亚硫酸氢盐转化提供了惊人的有效方法。随后可以通过许多不同的方法分析转化的DNA。

4. 发明授权

US07867701B2 Method for the simultaneous amplification of multiple sequences in a PCR reaction and marking thereof 失效
标题翻译：在PCR反应中同时扩增多个序列并进行标记的方法
公开(公告)号：US07867701B2
公开(公告)日：2011-01-11
申请号：US10451646
申请日：2001-12-22
申请人： Jürgen Distler , Kurt Berlin , Alexander Olek
发明人： Jürgen Distler , Kurt Berlin , Alexander Olek
IPC分类号： C12Q1/68 , C12P19/34 , C07H21/04
CPC分类号： C12Q1/686 , C12Q2537/143 , C12Q2525/155
摘要： A method is described for the amplification of nucleic acids, in which the segments to be amplified are first hybridized with at least two primer oligonucleotides that have two domains, wherein the sequence-specific domain found at the 3-end hybridizes to the segment to be amplified, while the generic domain found at the 5-end does not hybridize. Then an amplification reaction is conducted by means of a polymerase and subsequently a labeled primer oligonucleotide, which binds to the generic domain of the first primer, is hybridized to the amplificate which is formed. In the last step, the sequence of the amplificate is investigated.
摘要翻译：描述了用于扩增核酸的方法，其中待扩增的区段首先与具有两个结构域的至少两个引物寡核苷酸杂交，其中在3端发现的序列特异性结构域与所述区段杂交为而5端发现的通用域不杂交。然后通过聚合酶进行扩增反应，随后与第一引物的通用结构域结合的标记的引物寡核苷酸与所形成的扩增子杂交。在最后一步中，调查扩增序列。

5. 发明申请

US20090191548A1 METHODS AND NUCLEIC ACIDS FOR THE ANALYSIS OF GENE EXPRESSION ASSOCIATED WITH TISSUE CLASSIFICATION 审中-公开
标题翻译：用于组织分类相关基因表达分析的方法和核酸
公开(公告)号：US20090191548A1
公开(公告)日：2009-07-30
申请号：US12088384
申请日：2006-09-28
申请人： Kurt Berlin , Stephan Beck , Matthias Burger , Rene Cortese , Florian Eckhardt , Carolina Haefliger , Joern Lewin , Fabian Model , Alexander Olek
发明人： Kurt Berlin , Stephan Beck , Matthias Burger , Rene Cortese , Florian Eckhardt , Carolina Haefliger , Joern Lewin , Fabian Model , Alexander Olek
IPC分类号： C12Q1/68 , C07H21/04
CPC分类号： C12Q1/6881 , C12Q2600/158
摘要： The present application provides methods and nucleic acids the classification of a biological sample. This is achieved by the analysis of the expression status of at least one of the genes selected from Table 1 as disclosed.
摘要翻译：本申请提供了生物样品分类的方法和核酸。通过分析所公开的选自表1中的至少一种基因的表达状态来实现。

6. 发明申请

US20080064043A1 Method for detecting a methylation pattern 审中-公开
标题翻译：检测甲基化模式的方法
公开(公告)号：US20080064043A1
公开(公告)日：2008-03-13
申请号：US11881411
申请日：2007-07-26
申请人： Kurt Berlin , Juergen Distler , Reimo Tetzner
发明人： Kurt Berlin , Juergen Distler , Reimo Tetzner
IPC分类号： C12Q1/68 , C07H21/00 , G01N33/50
CPC分类号： C12Q1/6827 , Y10T436/143333 , C12Q2531/125 , C12Q2523/125 , C12Q2521/337
摘要： The present invention relates to a method for detecting the presence of one or more methylation patterns. It comprises the conversion of nucleic acids and a catalytic nucleic acid activity. Here, the conversion of the nucleic acid is characterized such that the 5-methylcytosine remains unchanged while the unmethylated cytosine is converted into uracil or another base, which can be distinguished from cytosine in its base-pairing behavior.
摘要翻译：本发明涉及一种用于检测一种或多种甲基化模式的存在的方法。它包括核酸的转化和催化核酸活性。这里，核酸的转化的特征在于5-甲基胞嘧啶保持不变，而未甲基化的胞嘧啶转化成尿嘧啶或其他碱，其可以与碱基配对行为中的胞嘧啶区分开。

7. 发明授权

US07229759B2 Highly sensitive method for the detection of cytosine methylation patterns 有权
标题翻译：用于检测胞嘧啶甲基化模式的高灵敏度方法
公开(公告)号：US07229759B2
公开(公告)日：2007-06-12
申请号：US10229370
申请日：2002-08-27
申请人： Alexander Olek , Kurt Berlin
发明人： Alexander Olek , Kurt Berlin
IPC分类号： C12Q1/68 , C12N15/11
CPC分类号： C12Q1/6827 , C12Q2523/125 , C12Q2537/163
摘要： The present invention concerns a method for the detection of cytosine methylation in DNA samples, wherein the following steps are conducted: (a) a genomic DNA sample, which comprises the DNA to be investigated and background DNA, is chemically treated in such a way that all of the unmethylated cytosine bases are converted to uracil, whereas the 5-methylcytosine bases remain unchanged; (b) the chemically treated DNA sample is amplified with the use of at least 1 primer oligonucleotide as well as a polymerase, whereby the DNA to be investigated is preferred as the template over the background DNA, and (c) the amplified products are analyzed and the methylation status in the DNA to be investigated is concluded from the presence of an amplified product and/or from the analysis of additional positions.
摘要翻译：本发明涉及检测DNA样品中胞嘧啶甲基化的方法，其中进行以下步骤：（a）将包含待研究的DNA和背景DNA的基因组DNA样品进行化学处理，使其所有未甲基化的胞嘧啶碱基被转化成尿嘧啶，而5-甲基胞嘧啶碱基保持不变; （b）使用至少1种引物寡核苷酸和聚合酶扩增化学处理的DNA样品，由此优选待研究的DNA作为背景DNA上的模板，（c）分析扩增产物并且待研究的DNA中的甲基化状态从扩增产物的存在和/或从额外位置的分析得出。

8. 发明授权

US07153671B2 Method for relative quantification of methylation of cytosine bases in DNA samples 失效
公开(公告)号：US07153671B2
公开(公告)日：2006-12-26
申请号：US10057776
申请日：2002-01-25
申请人： Kurt Berlin
发明人： Kurt Berlin
IPC分类号： C12P19/34 , C12Q1/68
CPC分类号： C12Q1/6827 , C12Q2565/137 , C12Q2563/107 , C12Q2531/113 , C12Q2523/125
摘要： A method is described for the relative quantification of the methylation of cytosine bases in DNA samples, wherein the following method steps are conducted: a) a genomic DNA sample is chemically converted with a reagent, wherein 5-methylcytosine and cytosine react differently and show a different base pairing behavior in the DNA duplex after the reaction; b) the DNA sample is amplified, whereby a fluorescently labeled dCTP or dGTP derivative is added; c) the amplified products are separated spatially from each other; and d) the fluorescence of the separated amplified products is measured quantitatively.

9. 发明申请

US20060204988A1 Method for determining the methylation pattern of a polynucleic acid 有权
标题翻译：确定多核酸甲基化模式的方法
公开(公告)号：US20060204988A1
公开(公告)日：2006-09-14
申请号：US11355417
申请日：2006-02-16
申请人： Anne Fassbender , Ralf Lesche , Juergen Distler , Christian Piepenbrock , Tamas Rujan , Kurt Berlin , Thomas Koenig
发明人： Anne Fassbender , Ralf Lesche , Juergen Distler , Christian Piepenbrock , Tamas Rujan , Kurt Berlin , Thomas Koenig
IPC分类号： C12Q1/68 , C12P19/34
CPC分类号： C12Q1/6827 , C12Q2531/113 , C12Q2525/191 , C12Q2521/331
摘要： Particular aspects relate to a method for determining the methylation pattern of a polynucleic acid, comprising: a) preparing a solution comprising a mixture of fragments of the polynucleic acid; b) coupling the fragments with a substance being detectable with a detection method; c) contacting a solution comprising the fragments of b) with a DNA microarray having a plurality of different immobilized oligonucleotides, each comprising at least one methylation site, at respectively assigned different locations thereon, the contacing under conditions affording hybridization of fragments with correlated immobilized oligonucleotides under defined stringency, and wherein the immobilized oligonucleotides have a length of less than 200 bases; d) optionally performing a a washing step; and e) detecting, using the physical detection method, such immobilized nucleic acids to which solution fragments are hybridized and/or to which solution fragments are not hybridized.
摘要翻译：具体方面涉及确定多核酸的甲基化模式的方法，其包括：a）制备包含所述多核酸的片段的混合物的溶液; b）将片段与可用检测方法检测的物质偶联; c）使包含b）的片段的溶液与分别具有多个不同的固定化寡核苷酸的DNA微阵列接触，每个寡核苷酸分别包含至少一个甲基化位点，分别在其上指定不同的位置，在提供相关的固定化寡核苷酸的片段杂交的条件下在限定的严格性下，并且其中所述固定的寡核苷酸具有小于200个碱基的长度; d）任选地执行洗涤步骤; 以及e）使用所述物理检测方法检测将所述固定化的核酸与所述溶液片段杂交并且/或所述溶液片段未杂交的固定化核酸。

10. 发明申请

US20050221314A1 Method and device for determination of tissue specificity of free floating dna in bodily fluids 审中-公开
标题翻译：用于测定体液中游离氨基酸的组织特异性的方法和装置
公开(公告)号：US20050221314A1
公开(公告)日：2005-10-06
申请号：US10506693
申请日：2003-03-05
申请人： Kurt Berlin , Andrzej Sledziewski
发明人： Kurt Berlin , Andrzej Sledziewski
IPC分类号： G01N33/50 , C12N15/09 , C12N15/10 , C12Q1/68 , G01N21/78 , G01N33/53 , G01N33/566
CPC分类号： C12N15/1003 , C12Q1/6806 , C12Q2600/156
摘要： The present invention relates to methods for detecting free floating nucleic acids, as present in not cellular bound nucleic acids in bodily fluids like plasma or serum fractions of human or animal blood or in any other tissue samples derived from the human or animal body in order to diagnose a cell proliferative disease. Specifically the invention relates to the detection of increased levels of nucleic acids in bodily fluids. Furthermore the invention allows to determine the source of the enriched DNA by measuring the ratio of DNA originating from a certain organ versus total DNA from other organs in a given bodily fluid sample by specifying the DNA's methylation pattern. This can be done with or without increasing the DNA concentration of a given biological sample. In a preferred embodiment a further analysis of this methylation pattern allows for the detection of the presence of tumourous or otherwise proliferative disease in said organ.
摘要翻译：本发明涉及用于检测游离漂浮核酸的方法，其存在于体液中的非细胞结合核酸中，如人或动物血液的血浆或血清部分或源自人或动物体的任何其他组织样品，以便诊断细胞增殖性疾病。具体地，本发明涉及检测体液中核酸的增加水平。此外，本发明允许通过指定DNA的甲基化模式，通过测量来自特定器官的DNA与给定体液样品中来自其他器官的总DNA的比例来确定富集DNA的来源。这可以在或不增加给定生物样品的DNA浓度的情况下进行。在优选的实施方案中，该甲基化模式的进一步分析允许检测所述器官中肿瘤或其他增殖性疾病的存在。

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