does anyone has any tips or a good protocol for the isolation of large quantities (>5 µg) of single copy plasmids (size=12.9 Kbp)? I tried different plasmid isolation kits (Zyppy, Machery and Nagel, ...) and most of the time it ended up in a smear. Maybe someone of you does it on a daily routine and can recommend a special protocol?
I know that people in former times did it using CsCl-gradients ...but if possible i would avoid that!
Another thing i read of is that mini-F plasmids tend to integrate into the genome ...anyone of you experienced that?
Any ideas or comments are appreciated!!!
Many thanks in advance!
Edited by pDNA, 18 December 2011 - 06:09 AM.