Development of a new in situ technique demonstrating methyaled cytosine by iso-thermal DNA amplification method

通过制作与染色体DNA上甲基化胞嘧啶以序列特异性方式特异性结合的ICON探针，通过锇反应形成强化学交联。通过所谓的PRINS方法，利用有界的ICON探针在特定染色体上建立了DNA延伸反应。在这个延伸茎中加入修饰的核苷酸，并通过传统的组织化学技术进行可视化。通过这种简单的方法，可以可视化特定染色体或DNA序列的甲基化状态。到目前为止，我们收集了很多基本的数据。我们目前正试图在小鼠睾丸精子发生过程中可视化单拷贝基因，并更新我们的协议，以获得更稳定和可靠的协议。

By making a ICON probe which specifically bound to methylated cytosine on chromosomal DNA in a sequence specific manner, a strong chemical cross-linking was made by osmium reaction. By so-called PRINS method, a DNA elongation reaction was established using bounded ICON probe on the specific chromosome. Modified nucleotides were incorporated during this elongation stem and visualized by a conventional histochemical techniques. By this easy, methyalation status of specific chromosome or DNA sequence could be visualized. So far we gathered a lot of basic and fundamental data. We are now currently trying to visualize a single-copy gene during spermatogenesis in mouse testis, and updating our protocol for more stable and reliable one.