While loading the equal amount of DNA in all wells for agarose gel electrophoresis, I am not getting the equal visualization of DNA in all lanes. Is it because EtBr binds with different affinities with intact DNA and fragmented DNA since I am performing experiments with fragmented DNA?
While visualizing on Geldoc, it seems that control well with intact DNA is being loaded with smaller amount of DNA, whereas I am loading equal amount in all the wells. Please suggest some possible reasons.
EtBr does indeed fluoresce less with fragmented DNA.
The position of your samples on the gel may account for differences (upper wells vs. lower wells, middle wells vs. lateral wells), be it with pre- or post-staining.
Many a times handling error results in conditions. The amount of sample to be loaded may vary depending on how deep of shallow you are loading the sample. Part of sample is lost with Shallow loading (at lesser depth) and amount thus actually loaded varies and results same as you observe.
- Badges
- Science topic
- Similar topics
- Gels