- Follow the instruction in Protoplasts Formation, Regeneration and Transformation
- Add 3.85 ml P buffer to 1 g PEG sterilised.
- Add 200 µl PEG-P buffer mixture and a DNA solution to 50 µl of the protoplast suspension.
- Plate it onto R5 or R2YE.
- Incubate the plate at 28-30ºC overnight (typically 12-18 hours).
- Overlay 1 ml sterilised water containing nalidixic acid and an appropriate antibiotic.