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Samples were reduced with DTT (5 L of 200 mM stock in H 2 O, 10 mM final concentration) and incubated at 65 C for 15 min, then alkylated with iodoacetamide (5 L of 400 mM stock in H 2 O, 20 mM final concentration) and shaken at 37 C for 30 min in the dark

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While results may develop more gradually than with some aggressive treatments, the improvements tend to be more sustainable and come without the risk of long-term damage or irritation that can accompany harsher approaches