I have purified a plasmid DNA using a plasmid miniprep kit for transforming into a bacterial expression system. I run an agarose gel for detection of the purified vector. I could detect three distinct bands for a purified plasmid DNA. Moreover, there was detectable smear. Does this imply degradation of the plasmid DNA?
No. You are most likely seeing the various coiled forms of an intact plasmid - positive supercoiled, relaxed, and negative supercoiled. Only linear DNA runs as a single band on a agarose gel. What you are seeing is totally normal.
No. You are most likely seeing the various coiled forms of an intact plasmid - positive supercoiled, relaxed, and negative supercoiled. Only linear DNA runs as a single band on a agarose gel. What you are seeing is totally normal.
http://bitesizebio.com/13524/how-to-identify-supercoils-nicks-and-circles-in-plasmid-preps/
This may help Alemu...
Dear Katie and Dear Abdul,
Many thanks for your answers. I appreciate it.
Cheers,
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