LARGE PLASMIDS

[lyons at wehi.edu.au]

I have had difficulty growing large (>10 kb) plasmids.  Others seem to be
able to get constructs of this size up.  Is this due to sequences 
differences, or are there tricks associated with growing or isolating
large plasmids.

Can anyone direct me bacterial strains that might be superior for this
purpose, or to plasmid backbones that might do it for me.  

What about growth conditions?  Is amplification likely to help? 
How about growing them cooler and slower?

Help!!

Thanks

Ian Lyons
The Walter and Eliza Hall Institute
Melbourne