Using this system for the 1st time.
(a)From what I read from other posts, many suggested retaining the ccdB gene but using a ccdB resistant strain will help in cloning of the control plasmid. Would you sugest any other plasmid?
(b)Assuming I am retaining the ccdB gene as my control gene, should I use the plasmid or different( eg pGWB5 for both gene of interest and control OR pGWB5 for gene of interest and pGWB20 for control)?
What do you need the control for, exactly? I'm not seeing much of an advantage to doing any of the things listed.
If it's to test competence of your E. coli, then cloning the control plasmid into a ccdB resistant strain wont tell you anything about the competence of the stock, since you'll end up using a different strain for cloning anyway.
If it's to make sure the enzymes are working, this isn't going to be informative for that.
Personally, when it comes to cloning I only really bother with a control if something isn't working. Otherwise it's kind of a waste of time.
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