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Alternatively produce a discontinuous gradient from 2 ml each the 27.6%, 20.7%, 13.8% and 4.6% Nycodenz solutions (see Step 2) by underlayering from a syringe and metal cannula and allow a continuous gradient to form by diffusion (see Notes 4 and 5)

This depletion can lead to a host of issues, including fatigue, skin concerns, and compromised immunity

Science 300(5619):644647