Protoplast Transformation: Difference between revisions
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*[[P buffer]] | *[[P buffer]] | ||
*1 g polyethylene glycol 1000 (PEG) | *1 g polyethylene glycol 1000 (PEG) | ||
*[[R5]] or [[R2YE]] plate | *[[R5]] or [[R2YE]] plate |
Revision as of 21:58, 24 March 2020
Materials
Instructions
- 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 298-30ºC overnight.
- Overlay 1 ml sterilised water containing nalidixic acid and an appropriate antibiotic.