Yep, I saw that, that's how I got the idea to try it :) Unfortunately haven't gotten the time to try it with PEG8000 yet
-- In my protocol I used JM109 cells without pKD46. It shouldn't be needed, and I don't know if expressing it helps.
-Koeng
On Wednesday, October 19, 2016 at 7:16:40 AM UTC-7, Cihan AYDIN wrote:
On Wednesday, October 19, 2016 at 7:16:40 AM UTC-7, Cihan AYDIN wrote:
Just learned about the plethora of information starting from Gibson assembly and came across this post.
If this is not you, someone have already tried the PEG8000 approach and pervailed :-) http://openwetware.org/wiki/SPLiCE
What I wonder is however this protocol also do not mention about inducing the pKD46 plasmid, is it expected (kind of D'uh! moment?) or it just leaks enough to make a difference (since it is araB, one should not expect leakage).
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