GSM6507444: cerebellar endothelial cells Ccm3 knock-out 1; Mus musculus; RNA-Seq
Endothelial cells were isolated from the cerebella of 3 wild-type and 3 Ccm3 knock-out mice with the Adult Brain Dissociation kit (130-107-677; MiltenyiBiotech), CD45 MicroBeads (30-052-301; MiltenyiBiotech) and CD31 MicroBeads (30-097-418; MiltenyiBiotech) according to the manufacturer's instructions. Total RNA was extracted from each sample with the RNeasy Micro Kit (Qiagen) in the automated QIACube (Qiagen) platform. RNA sequencing libraries were prepared with SMARTer Stranded Total RNA Sample Prep kit - Mammalian kit (Clontech), according to the manufacturer's instructions. This kit includes depletion of ribosomal RNA prior to library synthesis, barcoding and amplification. 1 μl of each library was analyzed in the Agilent 2100 bioanalyzer for validation of fragment size and quality control of the amplified RNA-seq library, using the High Sensitivity DNA kit (Agilent).