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DEAE transfections.

David L. Haviland, Ph.D. haviland at KIDS.WUSTL.EDU
Mon Dec 4 13:34:08 EST 1995


Greetings all:

We are attempting to do DEAE transfection for our reporter gene 
constructs.  I was wondering if anyone has any suggestions to get around, 
or deal with the toxicity of chloroquine used in the protocol.  
Apparently it is necessary to avoid acid damage of the transfected DNA 
but we seem to be incurring massive viability loss.  It is not uncommon 
for us to start with a 60% confluent plate and by the time the day ends, 
we're lucky to have 5% of the cells left.  We've already looked through 
Maniatis and the Harvard manual where the latter suggests using NU-serum 
to aid cell viability.  NU-serum is on indefinate back-order and we're 
trying alternatives.

Any thoughts?

Many thanks in advance.
David
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+  David L. Haviland, Ph.D.          .***.           .***.   .***.        +
+  Washington Univ. School of Med  * | | | *       * | | | * | | | *      +
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