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plasmid rearrangement or sabotage - (Oct/30/2012 )

A student in our lab made a AAV construct (with ITRs). He digested and confirmed that he got the right colonies. He put his glycerol stock (sure II cells) in -80C. A couple of days later he isolated plasmid and digested it. He found the plasmid is changed. I repeated his experiment and I got exact the same thing. So, do you think is this the plasmid rearrangement or sabotage?


The first digest- the confirmation one- how was that done? From a mini-prep? Colony PCR from the plate?


Thank you. pick up colony from plate and grow them up and isolate plasmid by mini-prep.