GSM9115603: Fipronil_biological_replciate_1; Danio rerio; RNA-Seq
For RNA isolation, sample tubes containing pools of 8 animals in 200 uL of 1X RNAshield were incubated at 32°C for 30 minutes to ensure complete thawing. 100 µL of 0.5 mm zirconium silicate beads were added, and samples were homogenized in the bullet blender at speed 10, for 5 minutes. Samples were then spun down for 5 minutes at 12000 xg, afterwards 250 µL of supernatant from each tube was aliquoted twice (for a total of 500 µL) into two new sample plates. These plates were then processed using the KingFisher Apex system for automated RNA extraction using Zymo Research's Quick RNA Magbead Kit and eluted into 50 µL RNAse/DNAse free water. The samples underwent PrepX Robotic RNA PolyA enrichment combined with NEBNext UltraII Directional RNA Library Prep/Illumina Stranded mRNA library preparation.