pZERO vectors: pros and cons?
gerdn at ibg.uit.no
Wed Jan 22 04:51:09 EST 1997
I have tried the pZero 2.1 (once); First of all; No system is
perfect: there is some background of colonies without insert, but
restreaking and comparing growth colonies showed that those without
insert grows badly, so with a little practice I think it should be
possible to pick the right colonies just by inspecting them carefully.
About which cells to grow them in - Why make up problems? - the best is
to start with the cells that comes with the kit , if you really need
another strain then you can worry aboutwhat have to be mutated.
compare descriptions of your strain and the one used in the kit
(TOP10F¥, i think it was)
How to grow the plasmid without killing the cells?: If you are using the
"default" strain - Just don¥t put IPTG in the medium.
Last: I got one hint (without asking for it) when I called the company:
Cut the plasmid as short time as possible!!!!
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