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METHOD FOR DETERMINING THE DEGREE OF METHYLATION OF DEFINED CYTOSINES IN GENOMIC DNA IN THE SEQUENCE CONTEXT OF 5'-CpG-3'

机译:测定5'-CpG-3'序列背景下基因组DNA中确定的胞嘧啶甲基化程度的方法

摘要

A method is described for the detection of the degree of methylation of a specific cytosine in the sequence context 5′-CpG-3′ of a genomic DNA sample. In the first step, the genomic DNA is chemically treated in such a way that the cytosine bases are converted to uracil, but not the 5-methylcytosine bases. Then segments of the genomic DNA which contain the said specific cytosine are amplified, whereby the amplified products are given a detectable label and in the following steps the extent of hybridization of the amplified products on two classes of oligonucleotides is determined by detection of the label of the amplified products, and a conclusion is made on the extent of methylation of said specific cytosine in the genomic DNA sample from the ratio of the labels detected on the two classes of oligonucleotides as a consequence of the hybridization.
机译:描述了一种在基因组DNA样品的序列背景5'-CpG-3'中检测特定胞嘧啶的甲基化程度的方法。第一步,对基因组DNA进行化学处理,以使胞嘧啶碱基转化为尿嘧啶,而不是5-甲基胞嘧啶碱基。然后,扩增包含所述特异性胞嘧啶的基因组DNA的片段,从而将扩增产物赋予可检测的标记,并且在随后的步骤中,通过检测标记的核苷酸来确定扩增产物在两类寡核苷酸上的杂交程度。通过扩增,从两类寡核苷酸上检测到的标记比例,得出基因组DNA样品中所述特异性胞嘧啶甲基化程度的结论。

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