title: GSM3712312: Mesothelium_4; Mus musculus; RNA-Seq
library_construction_protocol: 10 week old C57BL/6 mice were sacrificed.  The peritoneal cavities were washed by injecting 10 ml of 1x PBS (Sigma) via a syringe equipped with a 25G×5/8″ needle (BD). After gently shaking for 30 seconds, the fluid was removed and discarded. The peritoneum was filled with 5 ml of 0.25% Trypsin/EDTA solution (Gibco). The corpse was then maintained at 37°C for 20 min in a tissue-culture incubator before removal of the fluid. The detached cells were pelleted by centrifugation at 300×g for 10 min and resuspended in FACS buffer. The mesothelial cells were sorted from the cell suspension as CD45- PDPN+ GPM6a+ cells into DMEM + 10% FBS on a FACS Aria II flow cytometers (BD). Total RNA was extracted from the sorted mesothelial cells using the NucleoSpin RNA kit (Macherey-Nagel). rRNA was depleted using Ribo-Zero kit. RNA libraries were prepared for sequencing using standard Illumina protocols
attributes.tag: GEO Accession
attributes.value: GSM3712312