alias: GSM3267886
title: GSM3267886: P3 EphA4-GFP; Mus musculus; RNA-Seq
library_construction_protocol: Spiral ganglia were dissected and collected in Leibovit'z L-15 medium (Life technologies) on ice. Then the spiral ganglia (SG) were incubated in papain–DNAse solution (1.5 ml of papain at 1 mg/ml, 0.5 ml of DNAse at 0.1%) for 35 min (for P3 samples) or 25 min (for Adult samples) at 37 °C, shaking at 700 RPM. After spinning down the samples at 400 RCF for 10 min, the supernatant was removed and replaced by Dulbecco's modified Eagle's medium (DMEM) F-12 (Life technologies). SG were physically triturated using 3 different sizes of pipettes previously coated with 0.2% bovine serum albumin until the solution homogenized. The cell suspension was then filtered through 70 μm cell strainer (BD Biosciences) to remove clusters of cells. Single fluorescence positive cells were sorted by FAC sorting into individual wells of 384-well plates containing lysis buffer. Plates containing single cells were frozen immediately on dry ice and stored at -80 °C. We used 6 animals (12 cochleae) per experiment. RNA libraries were prepared according to Smart-seq2 protocol.
identifiers.namespace: GEO
identifiers.id: GSM3267886