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Isolation of a mini-F plasmid


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#1 pDNA

pDNA

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Posted 18 December 2011 - 05:30 AM

Dear people,

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!

Best regards,
p

Edited by pDNA, 18 December 2011 - 06:09 AM.





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