Mitochondrial DNA gene expression analysis by qPCR

Primary myoblasts were trypsinized and were re-suspended in 200 μL of PBS buffer containing Proteinase K (0.2mg/ml), SDS (0.2%), EDTA (5mM) for 3 hr at 56°C. After isopropanol precipitation, 50 ng of DNA was amplified by Q-PCR using specific primers for 12S, D-LOOP and BACT and Power SYBR® Green PCR Master Mix. Reactions were performed in triplicate and analyzed as described above with a normalization to BACT.