title: GSM8190742: Posterior segment, low light exposure, replicate 2; Convolutriloba longifissura; RNA-Seq
library_construction_protocol: Three replicates each containing five animals were used for RNA extraction for each condition. We rinsed the animals in ASW twice and then removed as much ASW as possible. 300 μl of Trizol was added, followed by a 2-3 min incubation at room temperature and vigorous vortexing to dissociate the tissue. Samples were then flash frozen on dry ice and kept at -80 °C until extraction so that all samples for each RNA-seq experiment could be processed in parallel. On the day of the extraction, samples were thawed on ice and 700 μl of fresh Trizol was added. After brief vortexing, 200 μl of chloroform was added and the sample was shaken vigorously for 15 s followed by a 2 min incubation. Samples were then centrifuged (16,000 g, 15 min at 4 °C) and the aqueous phase was carefully transferred into new tubes and processed with the Direct-zol RNA Purification Kit (Zymo Research, cat. #R2051) following the manufacturer's instructions, which includes a DNAse treatment step. RNA concentration and quality was quantified on a Bionanalyzer. Nextera XT library prep