title: HiSeq X Ten paired end sequencing; Transcriptomic tissue responses to pheromonal stimuli in Holothuria arguinensis
design: Transcriptomic tissue responses to pheromonal stimuli in Holothuria arguinensis
library_construction_protocol: Sea cucumbers were anesthetized by immersion in MgCl2 (5%) in sea water and tissues were dissected out (tentacles, nerve ring, and radial nerve) and immediately frozen on dry ice and kept at −80 °C for RNA extraction.  Total RNA was extracted from 40-45 mg of tissue using the E.Z.N.A. Total RNA Kit I (Omega Bio-Tek, USA) according to the manufacturer's instructions. Samples were treated with RNase-free DNase I (Omega Bio-Tek) to remove possible genomic DNA contamination. The integrity and quality of the extracted RNA was assessed by gel electrophoresis on a 0.8 % agarose TBE gel. The RNA purity and quantity were determined on a NanoDrop One (Thermofisher, Spain).  Sequencing library preparation and sequencing was conducted by Admera Health (New Jersey, USA) using a Illumina TrueSeq Stranded mRNA with Poly-A selection library kit (100- 500 ng RNA input)