title: GSM1669536: repo_micro_rep3; Mus musculus; RNA-Seq
library_construction_protocol: Mice were anesthetized and perfused with NaCl and organs were excised out. Brains were either fixed with 4% of PFA for histology, snap frozen in liquid nitrogen for RNA preparation, or digested according to manufacturer’s guidelines supplied in the MACS tissue dissociation kit Papain (Miltenyi Biotec). The tissue homogenate was loaded on a 30:70% Percoll gradient for enrichment of CNS infiltrates. The interphasic cellular ring was collected for further investigation. The different microglia samples were FACS sorted (FACS Aria) and immediately centrifuged down for 15 mins and 400 x g. RNA was isolated with the RNeasy® Plus Micro Kit (Qiagen) according to the manufacturer’s instructions. RNA amounts were measured by Q-bit 2.0 fluorometer (Life Technologies) and the quality was assessed on a Bioanalyzer (Agilent) using a High Sensitivity Chip. Samples with a RNA integrity number (RIN) > 8 were used for library preparation with TruSeq® RNA Sample Prep v2 Kit (Illumina) according to the manufacturer’s manual. Library size distribution was assessed on a Bioanalyzer 2100 using a High Sensitivity DNA Chip (Agilent). RNA libraries were prepared for sequencing using standard Illumina protocols
attributes.tag: GEO Accession
attributes.value: GSM1669536