I want to isolate to supercoiled plasmid from each other. I don't know is it possible by HPLC? and I don't know which column should I use? does anyone have such this experience?
It would be useful to know the size of your two plasmids, and what you intend to use the isolated DNA for.
I agree with Derek Brown, you can separate two plasmids of different size.
Hey there, an ion exchange column should work. Maybe this will serve as a guide DOI: 10.1016/j.jchromb.2007.04.005. I suggest using Tosho TSK-GEL DEAE column (Supelco) possibly a 0-1.0 M NaCl gradient and Tris buffer will do the trick.
Yes, If it is known the size of the plasmid, u can separate easily..
All scientist , Thanks for your answers.
I want to isolate a 2 kb plasmid from a 7 kb plasmid and Im searching for a suitable culumn. Because Ididnt such a experiment before.
Fabio, do you use this column?
Bhoj, whats the simple way for this aim?
Hi, indeed I have used this column for separating different topological forms of DNA with very good results and, more importantly, other researchers have use it as well. In the paper I suggested before you will find they use a TSKgel DNA-NPR column to separate 3.0, 5.5 and 7.6 kb plasmids; so yours should work too. However, no matter what the brand/make the column is, the consensus is that columns used for this application are anion exchage/ DEAE based.
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