GSM5269233: Lung_old_NaCl_mRNA_SK1.1.162; Mus musculus; RNA-Seq
Lung tissue was perfused twice with 10ml 1x PBS (Sigma, city, country) through the right side of the heart entering the pulmonary vasculature. Subsequently, lung was removed from the chest cavity, each lobe was separated and thereafter immediately flash frozen in liquid nitrogen. Lungs were then stored at -80°C until homogenized in 2ml Eppendorf tubes (Eppendorf, Hamburg, Germany) using a TissueLyser LT (Qiagen, Hilden, Germany). Samples remained frozen throughout the entire procedure. Lung powder was generated for next generation sequencing (NGS; 20mg tissue). Total RNA was isolated using the Qiazol procedure according to manufacturer´s protocol (Qiagen, Hilden, Germany). A total of 20mg tissue was used for RNA isolation. Quality of RNA was assessed by the Fragment Analyzer (Agilent Technologies, Santa Clara, CA, USA) using the SS RNA Kit (Agilent Technologies, Santa Clara, CA, USA).