RNA-seq of Hypomesus transpacificus: different tissues and life-stages : Sample Delta smelt_454sequencing
"Total RNA from multiple individuals exposed to a variety of stressors (i.e., temperature, salinity, hypoxia, turbidity, pH, exercise, olfactory, ammonia, copper, permethrin, esfenvalerate) were pooled to capture a diversity of mRNA transcripts related to the stress response in delta smelt. Total RNA was extracted from tissue samples (i.e., brain, heart, gill, kidney, liver, spleen, skin, muscle) or whole organisms using a combination of TRIzol Reagent (Life Technologies) and RNeasy kits (Qiagen), including on-column DNase digestion, as per manufacturer’s guidelines. RNA sample integrity was verified using a 2100 Bioanalyzer (Agilent Technologies) that indicated high quality RNA (RIN values >9). Two sequencing libraries were constructed from 5.0 ?g of pooled total RNA and libraries were sequenced using a Roche 454 GS-FLX Titanium system. Clean reads were assembled into contigs using Newbler software (Roche)."