Mice from the normative naive set (NON) and restraint/ethanol treatment sets (NOS, NOE, RSE and RSS) were killed at ∼2–3 months-of-age by cervical dislocation. Details on the strain, sex and age of each animal can be accessed from http://www.genenetwork.org/dbdoc/UT_ILM_BXD_hipp_NON_0909.html. Brains were removed quickly and stored in RNAlater (Ambion) overnight at 4 °C, then kept at −80 °C until use. The hippocampus was removed as described in Lu et al.,43 and left and right hippocampi were pooled and stored at −80 °C. RNA was isolated from these tissues using RNA STAT-60 (Tel-Test, www.tel-test.com) as per the manufacturer's instructions. RNA purity and concentration were evaluated with a spectrophotometer (NanoDrop Technologies, Rockland, DE, USA), and RNA integrity was assessed on the Agilent 2100 Bioanalyzer (Palo Alto, CA, USA). We required an RNA integrity number of >8. This RNA integrity number value is based on the intensity ratio and amplitude of 18s and 28s rRNA signals.
