MIDAS main protocol (aphidicolin 0.4 M) of HS68 cells (sequencing 2).
MIDAS main protocol. Thymidine-synchronized HS68 cells were released into S phase for 30 h with aphidicolin (0.4 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