What? LN2? Thats awesome but is there any specific reasoning for that? I've always just used a hot water bath and its just a bit of experimentation for the perfect exposure time since tube density, cell density, and uniformity of water in the bath may vary. Just streak on selection and compare colonies. My go to temp is 42 for 30sec. Average plasmid size is aboot 18k. Just try not to go over 42 unless you feel like trying it. It may be too much for the bug to handle. PEG for transforming bacteria seems overkill but by all means if you can get more efficiency then go for it. Do let us know if your peg method beats heatshocking.
As for peg, I'm not sure it can bridge peptidoglycan. Normally peg is an agent of aggregation so it can bring the plasmid and the bacteria closer but I cant see it working on its own. Do you have a ref for this? Normally peg is used as an alternative to agro and not as a means of transforming agro itself.
PS for your library I would recommend plant transformation technologies, the book. I think it would help you guys a lot since it covers particle and agro very well and is sourced from the labs that pioneered the tech. Its a bit pricey since its a Wiley Press product but well worth it.
Sebastian S Cocioba
CEO & Founder
New York Botanics, LLC
Sent via Mobile E-Mail
Ok, thanks.I looked up heat shock transformation of Agrobacterium, and just found the procedure using liquid nitrogen.So I assumed you can't do a normal heatshock. Therefore I plan using PEG transformation.For the toxin/antitoxin systems, do you know by chance a suitable promoter for E.Coli, that has good expression levels, but not super-high viral expression levels?--
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