GSM8606052: cleavage_mitoxantrone_rep1_nodrug_beta; Homo sapiens; OTHER
Sonication conditions were 5x10 second cycles at 20% power, 4μg and 5μg of TOP2A (3116) and TOP2B (3535) antibodies were incubated with protein A magnetic beads for 16 hours at 4°C and samples were eluted with 60μl TE buffer. Eluted DNA was treated with 8μl (1.12μg) of TDP2 (Abnova) for 40 minutes at 37°C to release DNA 5' ends that may be blocked for ligation by TOP2. DNA was digested with type 1 restriction enzyme BsmAI to optimise library construction. Samples were heat deactivated at 80°C for 10 minutes and DNA was purified using spin columns. Samples were eluted in 60μl TE buffer. Illumina Truseq