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    • 3. 发明申请
    • Method for detecting methylated CpG islands
    • 检测甲基化CpG岛的方法
    • US20060240460A1
    • 2006-10-26
    • US11398765
    • 2006-04-06
    • Gerd PfeiferTibor Rauch
    • Gerd PfeiferTibor Rauch
    • C12Q1/68C12P19/34
    • C12Q1/6813C12Q2537/164
    • 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.
    • 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。