rRNA depletion stranded TruSeq RNA-Seq of Orfelia fultoni (Appalachian Foxfire Fly), body without head and tail (see biosample for details), larva 8
In brief, a living Orfelia fultoni presumed last-instar larva (larva #8), was separated under a dissecting microscope using foreceps into "head", "body", and "tail" parts (see biosample metadata for further details) in a 34 mM sucrose solution. The respective part was placed into a separate 1.5 mL microcentrifuge tube, flash frozen in liquid N2, and placed in the -80C freezer. Later, 1 mL of Trizol was added to the tube. The larva part was next 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.