GSM5829422: DO_Mouse_High Fat_Muscle-96; Mus musculus; RNA-Seq
RNA was extracted from liver, muscle, and adipose samples collected from each animal at necropsy. Tissues (10 mg liver; 20 mg muscle; 60 mg adipose) were homogenized in the FastPrep 24-5G Homogenizer (MP Biomedicals, Irvine, CA) using Lysing Matrix D tubes. The RNA extractions were completed using the Maxwell RSC simplyRNA Tissue kit (Promega, Madison, WI).  Lysate pre-processing and cartridge preparation was done on the Promega Maxprep liquid handler and extractions were performed using the Promega Maxwell RSC48 instrument.  Sample concentrations were quantified  using the NanoDrop 8000 (Thermo Fisher Scientific) and RNA quality was checked on the Agilent 5200 Fragment Analyzer System (Agilent Technologies, Santa Clara, CA) using the Standard Sensitivity RNA kit (15NT). Samples with RQN ≥ 8 (muscle and liver) and RQN ≥ 7 (adipose) were used for sequencing. RNA libraries were prepared using TrueSeq Stranded Total RNA LP kit (Illumina, San Diego, CA) using manufacturer protocols and 100 ng of RNA. Sequencing was conducted using the MiSeq system  (Illumina, San Diego, CA).