Illumina HiSeq 4000 paired end sequencing; Gamma radiation induces Life stage-dependent reprotoxicity in Caenorhabditis elegans via impairment of spermatogenesis
Gamma radiation induces Life stage-dependent reprotoxicity in Caenorhabditis elegans via impairment of spermatogenesis
Worms were collected from NGS plates.  Worms were maintained for two months at 20 °C in swirling liquid cultures under dark condition (Brenner, 1974), in order to obtain a healthy stock population. Synchronous populations of nematodes were obtained by alkaline hypochlorite treatment as described by Stiernagle (2006) Gamma radiation exposures were conducted at the FIGARO experimental facility (Norderås, Norway) in the Norwegian University of Life Sciences (NMBU, Ås, Norway) (Lind et al., 2019). For every experiment performed in this study, synchronous cohorts of embryos or L1 nematodes were placed on NGM plates (Ø 3 or 6 cm) (1.7% agar, 2.5 mg∙mL-1 peptone, 25 mM NaCl, 50 mM KH2PO4 pH 6.0, 5 μg∙mL-1 cholesterol, 1 mM CaCl2, 1 mM MgSO4) with fresh Escherichia coli OP50 as a food source (cultured overnight at 37 °C in L-Broth medium, Lewis and Fleming (1995)). Experiments were conducted at 20 °C in the dark. For each experiment, three control NGM plates were placed behind lead shielding, and three plates per position were placed facing the front of the source at distances according to dose rates from 0.4 to 1490 mGy∙hr-1 RNA was extracted after 48 hours of exposure from L1 stage on L4-young adult nematodes (n=1000 per replicate) with Direct-zol Reagent (Nordic Biosite) and purified with RNeasy Mini Kit (Zymo Research) according to manufacture instruction. Strand-specific TruSeq™ RNA-seq paired-end libraries were prepared according to manufacture instruction.
Centre for Integrative Genetics (CIGENE), Department of Animal and Aquacultural Sciences (IHA), Faculty of Biosciences (BIOVIT), Norwegian University of Life Sciences (NMBU)
E-MTAB-8004:10 B 48h_p
Experimental Factor: irradiate
gamma
Experimental Factor: dose
10
Experimental Factor: time
48