curved gel band from mini prep

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alice.lunardon
alice.lunardon's picture
curved gel band from mini prep

 Hello,

I was trying to miniprep E.coli bateria to isolate a plasmid about 8kb long. When I check the miniprep product on gel I see a strange curved band on top of my presumed plasmid. Do you have any idea what it could be ad how I could obtain better results?

Thank you very much!!
AL

Brad Best
Brad Best's picture
I've never seen anything like

I've never seen anything like that.  Did you actually look at the gel on a UV box, or just take the image?  Those curved bands must have been blindingly bright.  They don't really look like they're running in the gel along with the DNA bands.  Almost like they're on top or something.

Chin Fen Teo
Chin Fen Teo's picture
Is it possible you overloaded

Is it possible you overloaded those lanes? Do you have any idea what is the concentration of your DNA?

alice.lunardon
alice.lunardon's picture
Thanks for the answers. I

Thanks for the answers. I loaded 2µL of miniprep on the gel and saw those intense bands also on the UV box. I did not check the concentration because I have RNA in my miniprep product. Could it be the effect of salts?

Chin Fen Teo
Chin Fen Teo's picture
I kind of think 2 ul of

I kind of think 2 ul of miniprep is a tons of DNA.... Can you try to make a 1/10 dilution and load like 1 or 2ul and see whether your gel resolution improves?

alice.lunardon
alice.lunardon's picture
I loaded 1µL of the 1:10

I loaded 1µL of the 1:10 dilution and now the gel the shows the expected plasmid bands without any curved bands, thank you for the suggestions!!