GSM9741434: Control_Day_2; Dascyllus aruanus; RNA-Seq
Total RNA was extracted from individual brain tissues using the Direct-zol RNA Miniprep kit (Zymo Research). Each brain was homogenized in 200 µL of TRIzol reagent (Invitrogen) using an electric pestle, cleared by centrifugation at 16,000 × g for 30 s at 4°C, mixed with ethanol, and processed per the manufacturer protocol including on-column DNase I digestion. RNA quality was assessed by Bioanalyzer (Agilent). TRUSEQ STRANDED samples (ED2TS, MD3TS, ML8TS) used for de novo transcriptome assembly: libraries prepared with the TruSeq Stranded mRNA Library Prep Kit (Illumina). INCPM samples (14 samples for DGE) used the INCPM mRNA-seq library preparation kit (non-stranded) at the Israel National Center for Personalized Medicine (Weizmann Institute). All libraries were sequenced on an Illumina NovaSeq 6000 with SP 300-cycle paired-end run (2 × 150 bp). The library was prepared with the addition 9 bp of Unique Molecular Identifiers (UMIs). UMIs are short sequences or molecular "tags" added to the DNA fragments in a sample library. These tags are added before PCR amplification and can be used to reduce errors and quantitative bias introduced by the amplification. they are listed as R2 in our fasta files