Protoplast Transformation: Difference between revisions
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==Materials== | |||
*[[P buffer]] | |||
*10.3% sucrose | |||
*1 g polyethylene glycol 1000 (PEG) | |||
*[[R5]] or [[R2YE]] plate | |||
==Instructions== | |||
#Follow the instruction in [[Protoplasts Formation, Regeneration and Transformation]] | #Follow the instruction in [[Protoplasts Formation, Regeneration and Transformation]] | ||
#Add 3.85 ml P buffer to 1 g PEG sterilised. | #Add 3.85 ml P buffer to 1 g PEG sterilised. | ||
Revision as of 21:57, 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.
