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The pellet was subsequently washed with 10 ml of the G-solution, which comprised of 500 ml HBSS (BE10-508F, Life Technologies) with 10% BSA (126615, Sigma Aldrich) and 1% Pen-Strep (15140122, Life Technologies), and re-suspended in 5.5 ml of gradient solution (15% Optiprep (5 ml 10% RPMI, Life Technologies) + 3 ml of 40% Optiprep, which was diluted from 60 % Optiprep with G-solution (D1556, Sigma Aldrich)) per sample, and placed on top of 2.5 ml of the gradient solution

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Getting it wrong does not mean disaster, but it can mean a rough 48 hours of nausea, vomiting, and gastrointestinal distress that makes you regret the decision

The good news is that these are usually mild and short-lived, often improving as your body adjusts