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

点击翻译按钮获取中文摘要

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.