RNA-Seq of Tripneustes gratilla: adult male neural
Whole RNA was extracted using a Qiagen RNeasy extraction kit. Extracts were treated with Epicentre Rnase-free Dnase I; 1 uL of Dnase I was added per 20uL of sample, then incubated at 37 C for 30 min. Samples were then cleaned using the Qiagen RNeasy Minelute Cleanup Kit. Quality was assessed on an Agilent 2100 Bioanalyzer. Poly-A tail isolation and cDNA synthesis were performed with the Illumina TruSeq Stranded mRNA Sample Preparation Kit (Protocol Part # 15031047 Rev. E) with no fragmentation time to allow for larger RNA fragments. cDNAs were sequenced on two lanes of an Illumina MiSeq with single strand chemistry for 300 cycles, using a V2 reagent kit.