title: GSM5421305: medaka108; Oryzias latipes; RNA-Seq
library_construction_protocol: RNA was isolated using a standard Trizol protocol, with minor modifications. Samples were thawed and vortexed for 10 seconds with 120 µl chloroform, incubated at room temperature for 3 minutes, and centrifuged at 14000g for 15 minutes at 4 °C. Glycoblue (Invitrogen) (17.5 µg/ml) was added to each tube to enhance precipitation and visualize the pellet. The aqueous phase (ca. 160 µl) was transferred to a new tube and RNA was precipitated in 160 µl 100% isopropanol by incubation at -20 °C for 1 hour, followed by centrifugation at 14000 g for 15 minutes at 4 °C. The pellet was then washed in 300 µl 75 % ice-cold ethanol and the sample centrifuged at 10000g for 10 minutes at 4 °C. The supernatant was removed and the RNA pellet briefly dried (approximately 5 minutes) at room temperature. The pellet was dissolved in 10 µl of RNase-free water. Samples were stored at -80 °C until use. cDNA was generated and amplified using the Smart-SEQ HT Kit for low RNA input according to the manufacturer's recommendations (Takara Bio), which was used for generating Illumina sequencing libraries with the Nextera XT kit.