Transformation advice needed.

David C. Rosen darosen at eis.calstate.edu
Fri Jul 29 14:19:58 EST 1994


I need to transform an INTACT plasmid of approximately 10kb.  I need a 
procedure that will reliably give me at least several transformants per 
plate.  The procedure will be repeated MANY times so I need it to be 
quite dependable.  
Questions-
1.  Will cells made competent with calcium chloride accomplish this?  
2.  What if the cells have been frozen after being made competent?  
3.  What if, after being thawed, the cells are held on ice for several 
hours before use?
4.  What other possibilities are there?  Electroporation is not a 
possibility and I do not want to buy competent cells.

Please send competence and transformation protocols.  Reply by e-mail
Thanks very much for your help.

David Rosen
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