公开(公告)号：US20060240460A1

公开(公告)日：2006-10-26

申请号：US11398765

申请日：2006-04-06

申请人： Gerd Pfeifer ,  Tibor Rauch

发明人： Gerd Pfeifer ,  Tibor Rauch

IPC分类号： C12Q1/68 ,  C12P19/34

CPC分类号： C12Q1/6813 ,  C12Q2537/164

摘要： The present invention provides new and improved assay for detection of genomic methylated CpG islands. This new method is termed the methylated-CpG island recovery assay (MIRA). In accordance with one embodiment, MIRA comprises the steps of: (a) incubating genomic DNA fragments with a methylated CpG island binding protein in the presence of a binding partner for the binding protein to produce bound DNA containing methylated CpG islands, (b) isolating the bound DNA, and (c) detecting CpG island methylation by gene-specific amplification reactions. In accordance with a preferred embodiment, MIRA comprises the steps of: (a) incubating sonicated genomic DNA with a matrix containing a fusion protein of glutathione S-transferase (GST) and MBD2b (GST-MBD2b) in the presence of MBD3L1 to produce bound DNA containing methylated CpG islands, (b) eluting bound DNA from the matrix, and (c) detecting CpG island methylation by gene-specific amplification reactions.

摘要翻译： 本发明提供用于检测基因组甲基化CpG岛的新的和改进的测定。 这种新方法被称为甲基化CpG岛回收测定（MIRA）。 根据一个实施方案，MIRA包括以下步骤：（a）在结合蛋白的结合配偶体存在下，将基因组DNA片段与甲基化CpG岛结合蛋白质孵育以产生含有甲基化CpG岛的结合DNA，（b）分离 结合DNA，和（c）通过基因特异性扩增反应检测CpG岛甲基化。 根据优选实施方案，MIRA包括以下步骤：（a）在MBD3L1存在下，将超声处理的基因组DNA与含有谷胱甘肽S-转移酶（GST）和MBD2b（GST-MBD2b）的融合蛋白的基质一起温育以产生结合 含有甲基化CpG岛的DNA，（b）从基质中洗脱结合的DNA，和（c）通过基因特异性扩增反应检测CpG岛甲基化。

公开(公告)号：US07425415B2

公开(公告)日：2008-09-16

申请号：US11398765

申请日：2006-04-06

申请人： Gerd P. Pfeifer ,  Tibor Rauch

发明人： Gerd P. Pfeifer ,  Tibor Rauch

IPC分类号： C12Q1/68 ,  C12P19/34

CPC分类号： C12Q1/6813 ,  C12Q2537/164

摘要： The present invention provides new and improved assay for detection of genomic methylated CpG islands. This new method is termed the methylated-CpG island recovery assay (MIRA). In accordance with one embodiment, MIRA comprises the steps of: (a) incubating genomic DNA fragments with a methylated CpG island binding protein in the presence of a binding partner for the binding protein to produce bound DNA containing methylated CpG islands, (b) isolating the bound DNA, and (c) detecting CpG island methylation by gene-specific amplification reactions. In accordance with a preferred embodiment, MIRA comprises the steps of: (a) incubating sonicated genomic DNA with a matrix containing a fusion protein of glutathione S-transferase (GST) and MBD2b (GST-MBD2b) in the presence of MBD3L1 to produce bound DNA containing methylated CpG islands, (b) eluting bound DNA from the matrix, and (c) detecting CpG island methylation by gene-specific amplification reactions.

摘要翻译： 本发明提供用于检测基因组甲基化CpG岛的新的和改进的测定。 这种新方法被称为甲基化CpG岛回收测定（MIRA）。 根据一个实施方案，MIRA包括以下步骤：（a）在结合蛋白的结合配偶体存在下，将基因组DNA片段与甲基化的CpG岛结合蛋白质孵育以产生含有甲基化CpG岛的结合DNA，（b） 结合DNA，和（c）通过基因特异性扩增反应检测CpG岛甲基化。 根据优选实施方案，MIRA包括以下步骤：（a）在MBD3L1存在下，将超声处理的基因组DNA与含有谷胱甘肽S-转移酶（GST）和MBD2b（GST-MBD2b）的融合蛋白的基质一起温育以产生结合 含有甲基化CpG岛的DNA，（b）从基质中洗脱结合的DNA，和（c）通过基因特异性扩增反应检测CpG岛甲基化。