plasmid digestion - problem with enzymes?pUC19
Posted 27 May 2013 - 04:53 PM
The enzymes I used are really old and I want to know if they are still working.
I've attached the photo of my gel - can anyone explain it?
Posted 28 May 2013 - 03:38 AM
An uncut plasmid preparation contains three forms, cut, nicked and uncut (this occurs spontaneously in a preparation) that all travel through the gel at different speed. Maybe your enzymes are too old and thats why only a fraction or none of have been cut and you see all three forms in your gel. Maybe you should try with new enzymes.
-sorry ignore comment it this was the case all three forms would appear in the same lane.
Edited by jmrebetz, 28 May 2013 - 04:08 AM.
Posted 28 May 2013 - 06:30 PM
Now I want to understand what has happend with the plasmid. Could it be the result of heating: the plasmid was not digested but it has changed it's conformation from circular to nicked.
Posted 09 June 2013 - 11:37 AM
Posted 12 June 2013 - 09:58 PM