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Table 1 Solutions for protoplast isolation and transfection

From: Simple protoplast isolation system for gene expression and protein interaction studies in pineapple (Ananas comosus L.)

Solution name Composition Storage Function
Enzyme solution 20 mM MES KOH, 1.5% (W/V) Cellulase R-10, 0.5% (W/V) Macerozyme R-10, 10 mM CaCl2, 20 mM KCl, 0.1% BSA (W/V) and 0.5 M Mannitol pH = 5.7 Freshly prepared Digestion of cell wall material
WS2 154 mM NaCl, 125 mM CaCl2, 5 mM KCl, 2 mM MES KOH pH = 5.7 4 °C Washing protoplasts
MMG 0.4 M Mannitol, 15 mM MgCl2, 4 mM MES pH 5.7 Freshly prepared Resuspend the protoplast for counting and transfection
PEG-Ca2+ solution 40% (W/V) PEG 4000, 0.2 M Mannitol and 0.1 M CaCl2 Freshly prepared Plasmid transformation
W1 0.5 M Mannitol, 20 mM KCL, 4 mM MES pH = 5.7 Freshly prepared Resuspend protoplast after transfection