Instrument: Illumina HiSeq 2000
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: SINGLE
Construction protocol: Entire brains were dissected from these four animals. Total RNA from these tissue samples was isolated as previously detailed using TRI-Reagent (Sigma Inc., St. Louis, MO, USA). Tissue samples were homogenized in TRI-Reagent followed by addition of 200 µl/ml chloroform and the samples vigorously shaken and subjected to centrifugation at 12,000 g for 5min at 4 ºC. Total RNA was further purified using RNeasy mini RNA isolation kit (Qiagen, Valencia, CA, USA). Residual DNA was eliminated by performing column DNase digestion at 25 oC for 15 min. Total RNA concentration was determined using a Qubit 2.0 fluorometer (Life Technologies, Grand Island, NY, USA). RNA quality was verified on an Agilent Bioanalyzer (Agilent Technologies, Santa Clara, CA) to confirm that RIN scores were above 8.0 prior to sequencing. RNA sequencing was performed upon libraries constructed using the Illumina TruSeq library preparation system (Illumina, Inc., San Diego, CA, USA)