Just an FYI: you can often substitute PEG3350 for PEG8000 in protocols. Just watch for molarity, substituting it gram-for-gram is probably unwise. Someone with a better grasp of how PEG affects osmolarity might chip in, here. :)
In cases where you *can* substitute, it's really handy, because PEG3350 is sold as an over-the-counter laxative under various brand names. In the US, it's known as Miralax, and I've successfully used it with other protocols that called for PEG3350.
On Wed, Oct 19, 2016 at 4:44 PM, Koeng <koeng101@gmail.com> wrote:
-- On Wed, Oct 19, 2016 at 4:44 PM, Koeng <koeng101@gmail.com> wrote:
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: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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