GSM6450588: eWAT_EX10970_20 [1060_0035]; Mus musculus; RNA-Seq
Total RNAs were extracted using the Ambion Magmax™-96 total RNA isolation kit (Applied Biosystems, Waltham, MA, USA) according to the manufacturer's instructions. As previously described43, 5 mg of tissue was placed in the lysis solution and homogenised in Tissuelyzer™ (Qiagen, Hilden, Germany) for 30 s. Nucleic acids were captured on magnetic beads, washed and treated with DNase. Total RNA was then eluted in 50 μl elution buffer. The RNA quality and concentration were measured using an RNA Pico chip on an Agilent Bioanalyzer (Agilent Technologies, Santa Clara, CA, USA). The cDNA libraries for RNA-Seq  were prepared using 1.2 ng total RNA input with the SMART-Seq v4 Ultra Low Input RNA Kit for RNA-Seq (Clontech Laboratories, Mountain View, CA, USA). RNA-Seq was performed as 100 bp single reads on a High Seq 2500 (Illumina, Inc., San Diego, CA, USA) as previously described.  For all RNA-Seq analyses, approximately 10–30 million reads per sample were obtained.