GSM3823779: MG-Sal-WT-4M; Mus musculus; RNA-Seq
Brains were finely minced with a razor blade and tissues were dissociated to single cell suspension with Accutase on ice for 30 min. Cell suspensins were resuspended with 33% Percoll and centrifuged for 15 min at 800g (no brake). Cells were washed, stained and subjected to sorting for CD11b(+)CD45(lo) cells. RNA was isolated from sorted microglia using Qiagen Universal AllPrep kit (Qiagen 80004) according to the manufacturer's instructions. cDNA for RNA-Seq libraries was prepared using 10 ng of RNA using the SmartSeq v4 Ultra Low RNA Input Library prep protocol (Clontech 634889) according to the manufacturer's instruction. The cDNA (250 ng starting material) was then tagged and fragmented with the NexteraXT DNA Library Preparation kit (Illumina FC-131-1024) according to the manufacturer's instructions. Quality of amplified cDNA and fragmented libraries was assessed on a Bioanalyzer with the High Sensitivity DNA chips (Agilent 5067-4627). Tagged libraries were pooled (8 nM total DNA) and sequenced on an Illumina HiSeq2500 sequencer, with target depth of 20 million of 50 bp paired end reads
GEO Accession
GSM3823779