Sequencing of RNA from C. fleckeri tentacle
Tentacles were excised and immediately preserved in RNAlater (Life Technologies). Total RNA was isolated from the C. fleckeri tentacles of a single specimen using a handheld rotor-stator homogenizer (QIAgen) and TRIzol (Life Technologies), according to the manufacturer’s instructions. The RNA was DNase-treated, purified using a Nucleospin RNA II kit (Macherey-Nagel) and eluted in nuclease-free water. RNA purity, concentration and integrity were evaluated using a NanoDrop 2000 UV-Vis spectrophotometer and an Agilent 2100 Bioanalyzer with a RNA 6000 Pico kit. Purified RNA (5 μg; RIN 9.1) was submitted to the Ramaciotti Centre for Genomics (Sydney, Australia) for RNA-seq library construction and 100 bp paired-end sequencing on an Illumina HiSeq2000 sequencer.