Do you find that PEG transformation works significantly better than electroporation on Arabidopsis protoplasts?
A3. I spent a lot of time trying to find the best electroporation condition for Arabidopsis protoplasts since I like it better than the PEG method (Machine is still more reliable than human hands and I was worried about the use of too much calcium needed for PEG transformation). With maize protoplasts, we can carry out 100 samples in 30 min with consistency. However, I don’t think I can really recommend anything that works well for Arabidopsis at the moment. I did find a condition that is described in the Arabidopsis mesophyll protoplast protocol. However, I have pretty much given up using it as a routine protocol since it’s not always easy to obtain reproducible results. For each cell type and each plant, you need to figure it out empirically and it takes a lot of time. I have not yet tried protoplasts isolated from culture lines. You may give it a try and it may turn out to be easier.
