GSM1207187: H10; Rattus norvegicus; RNA-Seq
Skeletal muscle (soleus) was collected from the hind limb of offspring and immediately snap frozen and stored at -80˚C for RNA and protein analysis. Frozen muscle (~50 mg) was homogenised with a motorized rotor-stator homogenizer (3 pulses of 15 seconds).  Total RNA was extracted using RNeasy isolation kit (QIAGEN, Valencia, CA) using the manufacturer’s recommended protocol. RNA quality was determined by the MultiNA capillary electrophoresis system (Shimadzu Biotech, Sydney, NSW, Australia). Messenger RNA was enriched using the NEBNext® Poly(A) mRNA Magnetic Isolation Module (New England Biolabs) from 2 μg total RNA. Sequencing libraries were prepared using the NEBNext® mRNA Library Prep Reagent Set for Illumina® (New England Biolabs). Libraries were quantified on MultiNA system and sequenced at a concentration of 12 pM on the Genome Analyzer IIx (Illumina Inc, CA, USA) using TruSeq version 5 cluster kit (cBot) and SBS kit (36 cycles).