rRNA depletion stranded TruSeq RNA-Seq of Orfelia fultoni (Appalachian Foxfire Fly), whole body, larva 6
In brief, a living glowing presumed last-instar Orfelia fultoni larva (larva #6), was transferred to a 2.0 mL microcentrifuge tube, 1 mL Trizol was added, and the tube was frozen at -80C. The next day, the tube was thawed and the larva was macerated with 1.5 L pipette tips, alternating with centrifugation at ~20,000 x g for ~15 seconds. This clarified Trizol solution was flowed through an Epoch Life Science EconoSpin RNA/DNA Micro Spin Column. The column was washed using typical silica-based RNA extraction buffers and procedures. The RNA was eluted off the column in 20 L EB buffer.