28,000-year-old mammoth cells have shown reactivity in mouse egg cells

Protein extraction from the mammoth tissues and following sample preparation were performed in a laboratory where modern elephantids have never been introduced. The skeletal muscle and bone marrow tissues (100 mg) were homogenised on ice in 300 μl lysis buffer comprising 7 M urea, 2 M thiourea, 4% (w/v) CHAPS, 0.05% (v/v) tri-n-butylphosphine and protease inhibitor cocktail (Roche Diagnostics) using Sample Grinding Kit (GE Healthcare). Four hundred μl of digestion buffer (300 mM TEAB containing 1 mM CaCl ) and 30 μl of sequence grade modified porcine trypsin (Roche Diagnostics) dissolved in the digestion buffer at 0.5 μg/μl were added to the samples, followed by incubation at 37 °C for 2 h. Subsequently, 50 μl of double distilled water and 30 μl of the trypsin solution were added to the samples, followed by incubation at 37 °C for 16 h.

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