申请人:——
公开号:US20040152080A1
公开(公告)日:2004-08-05
A method is described for the detection of 5-methylcytosine in genomic DNA samples. First, a genomic DNA from a DNA sample is chemically converted with a reagent, whereby 5-methylcytosine and cytosine react differently. Then the pretreated DNA is amplified with the use of a polymerase with primers of different sequence. In the next step, the amplified genomic DNA is hybridized to an oligonucleotide array and PCR products are obtained, which must be provided with a label. Alternatively, the PCR products can be extended in a primer extension reaction, wherein the extension products are also provided with a label.
In the last step, the extended oligonucleotides are investigated for the, presence of the label.
本文介绍了一种检测基因组 DNA 样品中 5-甲基胞嘧啶的方法。首先,用一种试剂对 DNA 样品中的基因组 DNA 进行化学转化,使 5-甲基胞嘧啶和胞嘧啶发生不同的反应。然后,使用聚合酶和不同序列的引物对经过预处理的 DNA 进行扩增。下一步,将扩增的基因组 DNA 与寡核苷酸阵列杂交,得到 PCR 产物,这些产物必须带有标记。另外,还可以在引物延伸反应中对 PCR 产物进行延伸,延伸产物也带有标记。
最后一步,对扩展后的寡核苷酸进行检测,以确定是否存在标签。