Dear all,
I'm currently working with halophilic archaea and I am trying to transform them using a spheroplasting protocol and PEG. The original protocols all use 60% PEG600, which unfortunately we do not have available at the moment (we will order it) and therefore I used PEG6000 because that was the lowest Mw available. Now the experiment did not work out, however I am not sure if this could be due to the PEG, I have been told that it does not matter that much. However, I would like to understand if the molecular weight of the PEG that you choose affect transformation efficiency? As for example in yeast I saw they use PEG6000 sometimes.
Thank you in advance!
If you read:
Cline, S.W.; Doolittle, W.F. Efficient transfection of the archaebacterium Halobacterium halobium. J. Bacteriol. 1987, 169, 1341-1344.
You will see in figure 3 there is a graph of PEG MW and transformation efficiency. It peaks at 600 and drops sharply after that. At 3350 it is near zero. Try to get the most pure preparation you can. Good luck.
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