MIDAS main protocol (aphidicolin 0.6 M) of U2OS cells.
MIDAS main protocol. Thymidine-synchronized U2OS cells were released into S phase for 16 h with aphidicolin (0.6 M) and RO3306 (9 M). G2/M-arrested cells were then washed and released for 1 h in medium containing nocodazole (200 ng/ml) and EdU (10M). HU (2 mM) was also added for the last 1.5h of the RO3306 treatment and for the entire time of the EdU and nocodazole treatment. Finally, mitotic cells were collected and the EdU-labelled DNA was isolated and sequenced