stability of plasmid

Wolfgang Schechinger Wolfgang.Schechinger at med.uni-tuebingen.de
Tue Aug 10 05:48:06 EST 1999


Hi Terry, 

Seems like DNAse to me (in any buffer or enzyme) -> check it 
this way: incubate aliquots of a plasmid you know that it's ok with 
the solutions to test. don't forger to test electrophoresis buffers.

Wo

> From:          Terry Lau <tcklau at yahoo.com>
> Subject:       stability of plasmid
> Date:          Tue, 10 Aug 1999 10:49:25 +0800
> Organization:  The Hong Kong Polytechnic University
> To:            methods at hgmp.mrc.ac.uk

> Hi,
> 
> Recently, I clone a insert(about 2.5k) by T/A cloning and I would
> like to cut the insert from the T/A vector. I inoculate the cell
> from the glycerol stock and grow in LB-AMP broth with shaking
> overnight. After miniprep and restriction digestion, I only obtain a
> small smear on the gel. Therefore, , I should retransform the
> plasmid every time. I don.t know what is the problem. Please give me
> any suggestion.
> 
> Thanks a lot!
> 
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Dr. Wolfgang Schechinger   
Pathobiochemistry Dept.      
University of Tuebingen, Germany
email: wgschech at med.uni-tuebingen.de * wwWait: http://www.medizin.uni-tuebingen.de/~wgschech/start.htm
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