I am trying to clone a gene of my interest into retroviral vector.
While maxipreping vector by transforming DH5a with vector DNA stock, I found strange debris in my liquid culture.
It looks like fibers floating in the culture, and when I centrifuge it aggregates with my bacterial pellet and don't resolve back.
I first thought it might be contamination and PCIAA + EtOH ppted the stock DNA. But the issue remains.
The vector is about 6.5kb long and has ampicillin resistant gene for selection.
Neither media or amp seem wrong since they worked perfectly in other cultures. I also maxipreped another clone which uses same vector but with a cloned DNA in it with no problem.
Thanks for reading and please get me some idea. I really need this cloned.
Edited by Wonyong, 26 March 2009 - 06:07 AM.