title: Cisplatin treatment
library_construction_protocol: Cisplatin (3 mg, 10 µmol) was activated by incubation with AgNO3 (6 mg, 35 µmol) at 37 °C for 6 hours in 50 µl of water. The activation reaction was conducted in a 1.5 ml Eppendorf tube covered with an aluminium foil to prevent light exposure. Cisplatin and AgNO3 mixture was centrifuged at 13,000 rpm for 10 minutes, and the clear supernatant containing activated cisplatin was used for DNA-adduct formation. Cisplatin-DNA adduct was formed by incubating DNA (10 µg) and activated cisplatin in 10 mM HEPES buffer, pH 7.0, at 37 °C for 1 hour in the dark. Cisplatin-treated DNA was purified by using Phenol-Chloroform extraction and eluted in 10 mM Tris, pH 8.0.                