title: Illumina MiSeq sequencing; RNA-seq experiment of isolated endothelial and microglial cells from hippocampal DG and CA1 regions under control and VEGF overexpression conditions
design: RNA-seq experiment of isolated endothelial and microglial cells from hippocampal DG and CA1 regions under control and VEGF overexpression conditions
library_name: Sample 5_s
library_construction_protocol: hippocampi were taken from Cx3cr1-GFP mice (3-4 mice pooled together per group). DG and CA1 were surgically removed under a stereoscope and collected into RNAse-free cold PBS. Dissociation was done using a neural tissue dissociation kit (Miltenyi Biotec, 130-092-628) and the gentleMACSTM Dissociator according to the manufacturer's instructions. Myelin was removed using myelin removal beads II (Miltenyi Biotec, 130-096-733). The same procedure was done for endothelial cell sorting using Tie2-GFP mice. Sorting was done using BD Aria FACS and only GFP positive cells were collected.  For endothelial cell sorting VEGF was switched-on (by tetracycline removal) for 1 months. For microglia sorting, VEGF was switched-on for 1 week. RNA was obtained using RNeasy plus Micro Kit (QIAGEN) according to manufacturer's guidelines The libraries were constructed using TrueSeq RNA V2 kit of Illumina http://www.illumina.com/products/truseq_rna_sample_prep_kit_v2.html.
attributes.tag: Experimental Factor: genotype
attributes.value: Tie2-GFP; Camk2a-tTa
attributes.tag: Experimental Factor: organism part
attributes.value: CA1 field of hippocampus
attributes.tag: Experimental Factor: cell type
attributes.value: endothelial cell