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plasmid rearrangement or sabotage


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#1 yqluuf

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Posted 30 October 2012 - 06:49 AM

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?

#2 leelee

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Posted 30 October 2012 - 06:26 PM

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

#3 yqluuf

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Posted 31 October 2012 - 05:57 AM

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




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