Hi everyone, I hope someone could help me. I have two bacterial strains that I isolated by streaking them in solid media and collecting an individual colony, repeating this process 2-3 times to ensure their purity. Then I extracted their DNA from a liquid culture and run an electrophoresis on Agarose gel 1%. Now, I found four bands, one of which I think may be a plasmid, but two of them are above my hypotetical genomic DNA. Is it possible for this bands to be a superplasmid? I also thought about the possibility of a contaminant, but I doubt it because all my media are superselective (for chimolitoautotrophs acidophiles) and also because I'm sure I cultured an isolated colony for each one.
Waiting for answers or hypothesis. I hope all of you are doing well :)
Running electrophoresis on genomic DNA will almost never give you single band.
Hello Jaime
What you have in your gel are isoforms of the genomic DNA. Yu have to consider small breakages in your genomic DNA during the extraction process. These small breaks would make genomic DNA to migrate at different rate due to more or less degree of coiling, thus you may see these bands.
Hey,
For genomic DNA extraction, follow the protocol perfectly. However, I'll suggest trying with 1.5/2% gel and observing the result. Because even if it's a breakage or super plasmid or mobile genetic elements, with thicker gel, you'll get to see the separation in high resolution.
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