RNA-seq of Patiria miniata: mature eggs
mRNA enrichment, cDNA synthesis, and Illumina sequencing was performed by Novogene Corporation. The mRNA from P. minata mature egg total RNA was purified with oligo(dT) beads. Next, the mRNA was fragmented randomly in a fragmentation buffer, then cDNA synthesis was performed with random hexamers and reverse transcriptase. Second-strand synthesis was performed after first-strand synthesis using a custom second-strand synthesis buffer and AMPure XP beads to purify the cDNA. The final cDNA library was prepared after purification, terminal repair, A-tailing, ligation of sequencing adapters, size selection, and PCR enrichment. The library concentration was first determined using a Qubit 2.0 fluorometer (Life Technologies), then the cDNA was diluted to 1ng/l and insert size was evaluated on an Agilent 2100 and the library was quantified to greater accuracy with quantitative PCR (library activity > 2nM).