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Digital PCR was performed using droplet digital PCR (ddPCR, BioRad Laboratories) for quantification of bottom:top strand ratios (Figs

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This confirms the pen mechanism is working and clears any air from the needle assembly

It does so in two ways (3): Altering substrate utilization during aerobic activity lasting more than 150 seconds Decreasing blood lactate accumulation with low-intensity aerobic activity, which may improve lactate threshold and ventilatory threshold 2