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Also, It started as 15 days

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The beads were washed with 15 mL of wash buffer (50 mM Tris pH 7.4, 300 mM NaCl, 0.5 mM DTT, 20 mM imidazole) and then with 5 mL elution buffer (50 mM Tris pH 7.4, 300 mM NaCl, 0.5 mM DTT, 500 mM imidazole)

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