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doi: 10.1016/j.jnutbio.2010.04.016

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)