title: C.elegans H2S RNA-seq time course
design: C.elegans H2S RNA-seq time course
library_construction_protocol: The animals were collected and rinsed three times with M9 buffer. After washing, the worm pellet was frozen in liquid nitrogen. To obtain RNA samples for RNA-seq, thirty young adult animals were picked to each fresh plate and allowed to lay eggs for 2 hours, after which the adult animals were removed and eggs were allowed to develop into day one adults.  Day one animals were subsequently challenged with H2S for three different time points (1 hour, 2 hours and 12 hours). For each time point, five plates of day one animals were exposed to 50ppm H2S, 150ppm H2S or 7% O2 as a control group. Animals were homogenized using Bullet Blender (Next Advance) in the presence of Qiazol Lysis Reagent and 0.5mm Zirconia beads at 4Â°C. RNA was prepared using RNeasy Plus Universal Mini Kit (Qiagen). Six independent RNA samples were prepared for all different treatments. The 2100 Bioanalyzer instrument (Agilent) was used for the RNA quality control with a microfluidic chip specific for RNA (Agilent RNA 6000 Nano Kit). 1 ug of qualified RNA samples in RNase-free ddH2O was used for library preparation. Library was constructed and sequenced by Novogene.