Field tested LOC for metaphase chromosome analysis

Field tested LOC for metaphase chromosome analysis UCPH will validate the chips developed in task 2.2 and 2.3 with relevant samples and bio-chemistry in order to develop methods and techniques to use the chips. Results from these experiments will be used for further improvements of the chip technology. Once we are able to handle the biological samples robustly in the chips we will start testing them on biologically relevant questions such as chromosome segregation dynamics. In this task we would aim to use purified proteins to address whether sister chromatid disjunction can be triggered solely by a combination of Separase and Topoisomerase II. We will also investigate the use of a modified version of cohesin incorporating a TEV protease recognition site into the SCC1 subunit. Using purified TEV protease, we could then trigger dissolution of cohesion ring and hence destroy sister chromatid cohesion. Our recent unpublished data indicate that the PICH protein strongly stimulates the catalytic activity of Topo II. Hence, we will investigate the possible role of PICH in stimulating centromeric DNA decatenation. The results of this study will be used in task 3.4 which aims at high resolution real-time imaging of the process.