I tried to isolate plasmid DNA from Agrobacterium tumefaciens using both manually and using kit and performed the electrophoresis on 0.5%, 0.8% and 1.0% agarose gel.
The ladder showed the bands on the gel.
(I didn't digest the plamids with a enzyme and the concentrations were not adjusted to the same)
The bands were smeared down, but no bands. I don't know why the bands smeared.
Or wasn't it the plasmid? kinds of different things like RNA, proteins, or chromosomal DNA fractions?
I really don't know why the bands were smeared if it was the plasmids.
I used the commercial plasmids DNA extraction kit from a company. And also follow the manual way.
Do you have any recommendation for this situation? or any advice?
Thank you.
Sincerely
It is very difficult to tell what is the problem from this gel, but based on the overall fluorescence, I would say you have vastly overloaded your lanes. Try using 1/10 and 1/50 the amount of plasmid and see what they look like. Then you might be able to see if you have DNA or only RNA or what.
yes, this is due to the over concentration of plasmid DNA. I further loaded the DNA in a low concentration and after gel run finally got the band. So thanks every one.
- Badges
- Science topic