GSM6939147: BulkHC2; Mus musculus; RNA-Seq
Dorsal hippocampi were dissected and immediately stored at -80°C in RNAlater solution. For RNA total extraction, hippocampi were homogenized in Qiazol using stainless steel beads. Chloroform was then added, and the homogenate was centrifuged at 12,000 x g at RT for 15 min. Aqueous phase containing RNA was precipitated using ethanol and then cleaned using the RNeasy kit. RNA was eluted in nuclease-free water, treated with DNase at RT for 25 min and precipitated in ethanol, sodium acetate (pH 5.2) and glycogen overnight at -20C. Precipitated RNA samples were centrifuged at top speed at RT for 20 min, washed with 70% ethanol and centrifuged at top speed for 5 min, dried and resuspended in nuclease free water. For bulk RNA-seq librairies were prepared using Ribo-Zero gold sample preparation kit. Library concentrations were measured using KAPA Illumina Library Quantification Kit. Polled libraries were sequenced on Illumina NovaSeq6000 sequencer with 150-bp Paired-End chemistry.