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Pinch a fold of skin at least 2 inches from the navel
The lysed tissue was centrifuged (13000 rpm, 3 min, 4 C) and the supernatant was transferred to a clean Eppendorf tube (low-bind protein) prior to protein concentration determination with the DC Protein Assay (Bio-Rad) according to manufacturers instruction
The benzyl alcohol preservative has been linked to severe toxicity in this population, including a potentially fatal condition known as gasping syndrome. Newborns have immature metabolic pathways and cannot efficiently metabolize benzyl alcohol, leading to its accumulation and metabolic acidosis with respiratory failure in severe cases
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