Hey,
I have a problem during my plasmid electrophoresis.
I got my plasmid band in the same line with 15kb marker. But when I digest it with a restriction enzyme I got my band in the correct line (6.2kb)
In a different plasmid isolation, same plasmid, I got both band at 15kb and 5kb( super coiled form).
Can this 15kb band refer for genomic DNA contamination?
I have added both photos. In the last isolation I also loaded some different samples so I cut and stick the plasmid lane and marker lane.
Thanks for answers.
Dear Enes...
If you are concerned about DNA contamination, a negative control sample will be helpful.
Kind Regards.
Looks too small to be gDNA, those bands tend to be close to the top of the gel. What bacterial strain are you using? Is it possible that there's another plasmid in there?
If you need to isolate the smaller plasmid, you could try gel extracting it and then transforming into new e. coli.
no worry, all are your right plasmid, single plasmid and plasmids ring chains. restrict will send all bands, bigs or smalls, to the right size.
image after I cut them with Xhoi and Mlui. I also cut them with EcorV both isolations gave the same band profile at 6kb band ( marker isnt seen good on photo).
Thanks for answers.
You need to run out your samples on your gel much longer. The ladder is too compressed to get an accurate size estimate.
problem is due to photo. It was able to be discriminated by eye directly, without photo.
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