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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
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。
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公开(公告)号：US20080220418A1
公开(公告)日：2008-09-11
申请号：US11664367
申请日：2005-09-30
申请人： Matthias Ballhause , Kurt Berlin , Dimo Dietrich , Antje Lukas , Matthias Schuster , Ute Wagner , Reinhold Wasserkort , Heike Ziebarth
发明人： Matthias Ballhause , Kurt Berlin , Dimo Dietrich , Antje Lukas , Matthias Schuster , Ute Wagner , Reinhold Wasserkort , Heike Ziebarth
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, an 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 。 这些方法是容易的，具有成本效益的，其特征在于重现性和可靠性高。 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。
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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
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。
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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.
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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
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。
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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。
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公开(公告)号：US07407749B2
公开(公告)日：2008-08-05
申请号：US10416624
申请日：2002-10-25
申请人： Kurt Berlin , Matthias Ballhause , David Gütig
发明人： Kurt Berlin , Matthias Ballhause , David Gütig
CPC分类号： C12Q1/6827 , C12Q2565/518 , C12Q2531/113 , C12Q2523/125
摘要： A method is described for the analysis of cytosine methylation patterns in genomic DNA samples. In the first method step, the genomic DNA is isolated from cells or other accompanying materials and bound essentially irreversibly to a surface. Then the DNA bound to the surface is treated, preferably with a bisulfite, in such a way that cytosine is converted into a base that is different in its base pairing behavior in the DNA duplex, while 5-methylcytosine remains unchanged. Then the reagents that were used are removed in a washing step. Finally, selected segments of the immobilized DNA are amplified in a polymerase reaction and the amplified products are investigated with respect to their sequence.
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8.发明申请Method for the detection of cytosine methylations in immobilized DNA samples 有权检测固定化DNA样品中胞嘧啶甲基化的方法公开(公告)号：US20080113379A1
公开(公告)日：2008-05-15
申请号：US11981357
申请日：2007-10-31
申请人： Kurt Berlin , Matthias Ballhause , David Gutig
发明人： Kurt Berlin , Matthias Ballhause , David Gutig
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6827 , C12Q2565/518 , C12Q2531/113 , C12Q2523/125
摘要： A method is described for the analysis of cytosine methylation patterns in genomic DNA samples. In the first method step, the genomic DNA is isolated from cells or other accompanying materials and bound essentially irreversibly to a surface. Then the DNA bound to the surface is treated, preferably with a bisulfite, in such a way that cytosine is converted into a base that is different in its base pairing behavior in the DNA duplex, while 5-methylcytosine remains unchanged. Then the reagents that were used are removed in a washing step. Finally, selected segments of the immobilized DNA are amplified in a polymerase reaction and the amplified products are investigated with respect to their sequence.
摘要翻译： 描述了用于分析基因组DNA样品中胞嘧啶甲基化模式的方法。 在第一种方法步骤中，基因组DNA从细胞或其他伴生材料中分离出来，并且基本上不可逆地结合到表面。 然后，优选用亚硫酸氢盐处理与表面结合的DNA，使得胞嘧啶转化为DNA双链体中其碱基配对行为不同的碱基，而5-甲基胞嘧啶保持不变。 然后在洗涤步骤中除去所用的试剂。 最后，在聚合酶反应中扩增所选择的固定化DNA段，并根据其序列研究扩增产物。
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公开(公告)号：US20060134643A1
公开(公告)日：2006-06-22
申请号：US10964167
申请日：2004-10-12
申请人： Kurt Berlin , Matthias Ballhause , Karen Cardon
发明人： Kurt Berlin , Matthias Ballhause , Karen Cardon
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。
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公开(公告)号：US08257950B2
公开(公告)日：2012-09-04
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
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。
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