title: GSM2310340: Microglia control (Sample 1); Danio rerio; RNA-Seq
library_construction_protocol: Zebrafish were sacrificed with ice water after 24 or 48 hours of treatment with MTZ (activated microglia) or DMSO (control microglia). Then brains were taken out of the skull and trypsinized on ice for 2 hours. Percoll gradient centrifugation was used to enrich the cell population for microglia. FACS sorting based on GFP expressing was used to isolate microglia. To extract the other brain cells, the GFP-negative population was collected. Cells were spun down and pellet was flash frozen in liquid nitrogen. Total RNA extraction was performed using the Qiagen miRNeasy kit. cDNA libraries were created using the SMARTer Ultra Low Input RNA Kit for Sequencing - v3 (Clontech). Illumina RNAseq libraries were prepared from cDNA using the Illumina TruSeq™ RNA Sample Prep Kit v2 according to the manufacturer’s instructions (Illumina Inc.). In all libraries 50 nucleotide single-end reads (SR50) were sequenced on an Illumina HiSeq2500 sequencer according to the manufacturer’s protocol, obtaining 10-20 million reads per sample library.
attributes.tag: GEO Accession
attributes.value: GSM2310340