title: Illumina HiSeq 4000 sequencing; Mouse tissue atlas
design: Mouse tissue atlas
library_construction_protocol: C57BL/6N mice (16 weeks old, m/f) were sacrificed individually by CO2 asphyxiation. Afterward, animal were perfused with 50 mL of precooled phosphate buffered saline, supplemented with cOmplete EDTA-free protease inhibitor cocktail (Roche) and a mixture of phosphatase inhibitor cocktail 1, 2 and 3 (Sigma-Aldrich). Tissues and organs were dissected, snap-frozen in liquid nitrogen and stored at -80 Â°C. Frozen tissues were then pulverized on liquid nitrogen. cDNA libraries were prepared using the TruSeq Stranded mRNA Sample Preparation kit (Ilumina) according to the manufacturer instructions Total RNA was isolated using the RNeasy Mini Plus Kit (Qiagen), according to the manufacturer instructions.  RNA was quantified (Nanodrop, Thermo Fisher Scientific) and quality checked with a Tape Station D1000 (Agilent Technologies). RIN values between 7 and 10 were accepted for further analysis.