Dose-Dependent Genomic Analyses in C. elegans

L4 stage animals were irradiated with doses of 20 Gy, 40 Gy, 60 Gy and 80 Gy using a Cs-137 source (IBL 437C, CIS Bio International), and doses of 50 J, 100 J, 200 J and 500 J using a Waldman UV 236 B device with a spectrum of 280 nm to 360 nm (UV-B / UV-A light). cisplatin exposure was performed as described [27 (link)]. In short, animals were incubated under gentle shaking for 16h at 20°C in M9 liquid culture with cisplatin (Sigma-Aldrich, P4394) concentrations from 0 to 400 μM (S1 Table). Treated animals were allowed to recover and 3 times 3 adults per genotype and dose were transferred onto fresh plates 24h post treatment. Adults were removed after 4h and eggs laid within this time were allowed to hatch and scored for viability [26 (link), 27 (link)]. 2 F1 L4s per plate were then singled. For higher doses of radiation, not all F1 animals produced progeny. Progeny of only one of each set of 2 F1 lines was frozen, providing 3 independent lines per genotype and treatment. Genomic DNA was isolated using Invitrogen ChargeSwitch® gDNA Mini Tissue Kit (Thermo Fisher Scientific, CS11204) and sent for sequencing.