title: GSM5471492: Mesenchymal Irradiation 2; Mus musculus; RNA-Seq
library_construction_protocol: Lungs were minced manually and then digested for 30 min at 37 degrees in a HBSS digestion solution containing DNase I (Sigma-Aldrich) and Liberase TM and TH (Roche Diagnostics). Samples were then washed, passed through a 100 um filter and incubated with Red Blood Cell Lysis buffer (Miltenyi Biotec) for 3-5 min. After a wash with MACS buffer (0.5% BSA and 250 mM EDTA in PBS), samples were passed through a 40 um filter and a 20 um strainer-capped flow cytometry tube to generate a single cell suspension. Samples were stained with a CD45, CD31, Ter119 and an EpCAM antibody.  CD45-CD45-Ter119-EpCAM+ and D45-CD45-Ter119-EpCAM- cells were isolated via FACS. Total RNA was isolated using the miRNeasy Micro Kit (Qiagen, cat# 217084), optimised for high quality RNA extraction from samples with low cell numbers, according to the manufacturer's instructions. Library generation was performed using the KAPA RNA HyperPrep with RiboErase (Roche).