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Need help with interestingdeletion problem - (Sep/06/2001 )

My entire lab is having a very bizzare problem. We are trying to subclonea insert into a plasmid. Minipreps show either a self ligated plasmid of the proper size or a plasmid less than half the proper size with theproper size insert. We have tried everything we can think of (ie. changing hosts, changing restriction site) but always have the sameproblem.

Any suggestions would be greatly appreciated.


Interesting problem. It looks like your lab has environmental aeromonas contamination which is clipping your target DNA with a site specific endonuclease after isolation. Leave an LB agar plate open on the benchtop overnight and you will see the bug. Use sterilized water and do all buffer transfers in a Bacteriological Hood.

Good Luck