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Protoplasts from barley endosperm A pellet of crude protoplast, obtained at 40-50 g for 2-4 min, was suspended in 2 ml of 40% (w/v) Nycodenz (in 50 mM CaCl2, 25 mM MES, pH 5.5), overlaid with 1 ml each of 26.6%, 13.3% and 6.6% Nycodenz (produced by dilution of the 40% Nycodenz solution with the protoplast medium)

This equalizes the pressure, making the water addition smooth and controlled

J Mol Endocrinol (1993) 10(3):25968