CZV8-top agar 100 mL per transformation 200 mLCzapek-Dox liquid 4.54 g 9.08 gagar 0.75 g 1.5 gsorbitol 18.22 g 36.44 gcentrifuged V8 juice 20 mL 40 mLadd H2O to 100 mL 200 mLadjust to pH 6.0 with NaOH split batch in 2 (½ for plating protoplasts, ½ for overlay)autoclave