seizure_seq
RNA was extracted from blood samples (stored in Paxgene tubes at -20 °C) using the Pre-Analytix RNA extraction kit (Qiagen). RNA-seq libraries were constructed using the Ion Total RNA-Seq Kit v2 (ThermoFisher Scientific) with 500 ng total RNA as starting material (not globin depleted). RNA was sheared, ligated to adapters, and reverse transcribed with Super ScriptTMIII. cDNA was size selected using Ampure XL beads (Beckman Coulter) and amplified using PlatinumTM DNA polymerase (15 cycles) with IonXpress barcode primers (1:16). Libraries were quantified using High Sensitivity DNA Bioanalyzer chips, pooled and cloned onto sequencing spheres, loaded on Ion 540 chips, and sequenced (Ion Torrent S5).