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Briefly, buffers provided with the kit were supplemented with Protease Inhibitor Cocktail (P8340, Sigma) and used to lyse the cells with a Dounce homogenizer, followed by separation by variable speed centrifugation

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A report from the Institute of Medicine released in March 2015 predicted that raising the smoking age to 21 nationally would reduce the amount of new smokers aged 15-17 by 25 percent

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