Sequencing of enriched mitochondrial DNA: human skeletal muscle tissue,A467T|W748S variant POLG: Sample M30A
Linear DNA, predominantly nuclear, was digested with Exonuclease V and a cocktail of seven restriction endonucleases that did not cut mtDNA. Remaining DNA was column purified, concentrated and the nuclear and mtDNA components quantified via real-time PCR. If the mtDNA was found to be under 30% of the total, then nuclease treatment and purification steps were repeated. Sequencing libraries were prepared via transposon-mediated tagmentation using the minimum sufficient number of PCR amplification cycles.