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Then, these samples were lysed in cell lysis/wash buffer (150 mM NaCl, 5 mM EDTA [pH = 7.5 at 25 C], 50 mM Tris-HCl [pH 7.5], 0.5% NP-40) containing protease inhibitor for 10 min on ice and then sonicated to generate DNA fragmentation with length of 200500 bp in shearing buffer (1% SDS, 10 mM EDTA [pH = 8.0], 50 mM Tris-HCl [pH = 8.0 at 25 C]) containing protease inhibitor using a Diagenode BioruptorPlus sonicator (30 s on and 30 s off for 12 cycles)

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Protein intake adequacy (1.2-1.5g/kg ideal body weight) + resistance training aren't optional add-ons

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The collagen mechanism is copper-dependent