Instrument: AB SOLiD 4 System
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: SINGLE
Construction protocol: Tissue from dorsal lateral prefrontal cortex (DLPFC) was pulverized over dry ice. Approximately 300 mg was weighed while frozen and stored at 80°C until extraction. Total RNA was extracted with TRIzol Reagent (Invitrogen Life Science, Cat. No. 15596-018, Mount Waverley, Victoria), using a polytron. Total RNA pellets were re-suspended in DEPC treated water. The yield of total RNA was then analysed using a spectrophotometer (Nanodrop ND-1000, Thermo Scientific). Total RNA was not further purified through a column. The quality of extracted total RNA was determined by high resolution capillary electrophoresis using the Agilent Bioanalyzer 2100. Twelve micrograms of RNA from each of the samples was used in creation of the library for sequencing. RNA was DNase treated using TURBO DNA-free (Ambion, Austin TX, USA) before the depletion of ribosomal RNA using the Ribominus kit (Invitrogen, Carlsbad, CA, USA). The resulting RNA was fragmented and the SOLiD library prepared using the Total RNA-Seq kit (Invitrogen). Quality control of the library was assessed by size (480% and within 200--300 bp) using a bioanalyser. Samples were run with four individuals per slide on the SOLiDv4 (Life Technologies, Carlsbad, CA, USA). Reads were unpaired and 50nt in length. Average reads per individual were 135 355 990 (±32 081 820) and did not differ between diagnostic groups.