Integrative genomic and transcriptomic analyses identify functionally relevant causal genes for exercise in a large animal model
Integrative genomic and transcriptomic analyses identify functionally relevant causal genes for exercise in a large animal model
Percutaneous needle muscle biopsies (approx. 300 mg) were obtained from the ventral compartment of the middle gluteal muscle from standing unsedated horses using a previously described method [53] and preserved in RNAlater (Thermo Fisher, Massachusetts, United States). Total RNA was extracted from approximately 70 mg tissue, using a protocol combining TRIzol reagent (Thermo Fisher, Massachusetts, United States), DNase treatment (RNase free DNase) (Qiagen, Hilden, Germany) and RNeasy Mini-Kit (Qiagen).RNA was quantified using a Nano Drop ND1000 spectrophotometer V 3.5.27 and RNA quality and purity were assessed using the 18S/28S ratio and RNA integrity number (RIN) on an Agilent Bioanalyser with the RNA 6000 Nano LabChip kit6 (Agilent, Cork, Ireland). Indexed, strand specific Illumina sequencing libraries were prepared using the TruSeq Stranded mRNA Library Preparation Kit LT (Illumina, San Diego, United States).