I have obtained some plasmids used as integration vectors, this question may apply to all plasmids. I would like to have a somewhat continuos source for these plasmids, let's say that the origin of replication is e-coli specific and can i then just transform the e-coli with the plasmid, let it grow to some density and then do miniprep? or can i also create glycerol stocks out of the bacteria that contain the plasmids and then harvest more plasmids as needed? i realize that both things are possible, but is this a good reasoning to follow?



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What I would usually do is transform E. coli with the plasmid, grow an overnight culture and mini prep the plasmid in the morning. Before you do the miniprep save a glycerol stock. Thus, you would have some plasmid to work with in the next couple of weeks, and also a glycerol stock to come back to. You can also keep the plate with your transformants, as you can go back to it within a month.

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    $\begingroup$ I'll add to this as well. Always keep a small aliquot of your miniprep DNA in a stock box, and keep it frozen, just in case the glycerol stock fails for some reason (e.g., freezer failure). The DNA is more forgiving of freeze-thaws and offers one more backup. I only do this with final (or destination) vectors that I have finished manipulating. $\endgroup$
    – user560
    Mar 5, 2014 at 22:35
  • $\begingroup$ Yeah, I think this is a good backup. $\endgroup$ Mar 5, 2014 at 22:37

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