GSM4861440: PS19_HA rep2; Mus musculus; RNA-Seq
hippocampi were dissected and snap frozen with liquid nitrogen in 1mL Trizol, and stored at -80°C. For RNA isolation, samples were thawed at 37°C then kept on ice prior to homogenization with Polytron homogenizer with a 5 second pulse. After a 5 minute incubation at room temperature, one tenth the sample volume of 1-bromo-3chloropropane was added, mixed by inversion and incubated for 3 minutes at room temperature. Samples were then centrifuged at 13,000 X g for 15 minutes at 4°C to separate the aqueous and organic layers. As much of the aqueous layer was recovered as possible, then RNA was precipitated with isoproponal. Pellets were then washed with 75% ethanol and resuspended in 50μL of dionized water. RNA library preparation and sequencing were performed by HudsonAlpha Genomic Services Lab. RNA was Poly(A) selected and 300bp size selected