Please can anyone suggest, weather DMSO is added (as a cryoprotectant) or not, to fungal protoplast. Since DMSO is a polar aprotic solvent I am scared of getting the protoplast membrane dissolved. If not, please suggest any other method. Thank you in advance.
For plasmid DNA transformation by PEG/elctroporation mediated methods.
Thank you
Hi Challagandla,
When I did PEG-mediated Arabidopsis and tobacco protoplast transformation (of plasmids), we did not use DMSO. But, I don't know whether DMSO is included in the fungal protoplast transformation protocol. I have seen some fungal electroporation-mediated protoplast transformation online. Have you checked out one of these papers?
Dear Yuan,
I gone through few related papers, but I didn't find any one using DMSO. But in a Plant Fungal Interaction Protocols- 2012 (chapter-16) they have mentioned to add 7% DMSO.
Thank you
Hi Challagandla,
Since both with DMSO and without DMSO (several papers) were used in protocols and succeed, you can divide a batch of protoplasts into two parts for experiments, one adding DMSO and the other without adding DMSO for comparison. Then, you can find out which one is better in person. Otherwise, since several papers already published with successful results without adding DMSO, I would pass it.
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