GSM6559814: Octopus Adult Arm Left 1 Tip [OB2]; Octopus bimaculoides; RNA-Seq
For each sample, tissues stored at -80°C were ground in liquid nitrogen using a mortar placed in dry ice. Resulting powder was aliquoted and 15 mg of this powder was used to extract  RNA. RNA was extracted using Trizol (Invitrogen) following the manufacturer's instructions with some modifications. Briefly, during precipitation in isopropanol, 10 µg of Glycoblue (Thermo Fisher Scientific) was added and precipitation was performed overnight at -20°C followed by 1 hour centrifuge at 12000g at 4°C. RNA was resuspended in water . RNA was treated by DNAse I for 30 minutes at 37 °C followed by RNA purification (RapidOut DNA Removal Kit – Thermo Fisher Scientific). RiboZero was used to remove ribosomal RNA and the remaining sample was used for library preparation according to manufacturer's instructions (Illumina) from 250 ng of RNA. Libraries were sequenced on NextSeq550 (Illumina) to obtain 150 bp paired end reads.