2% agarose gel staine dwith ethidium bromide. I'm not sure why there is irregular staining but it make sit difficult to see the bands I'm looking for. The bands should be at 369 bp and you can faintly see the positive control, so these weird patterns are making it hard to see if any of the other samples have the gene I'm looking for.
new 1x TBE buffer was used. Ethidium bromide was added when the agarose was at 55 celcius
Thanks!
Hello Velic,
What was the final concentration of EtBr that you added in the gel?
Generally, a concentration of 0.5 microgram/ml will give optimum results. Try adding some EtBr in the running buffer to prevent migration of EtBr from your gel (as visible from lower part of your gel).
-Bhuwan
this effect looks like the gel was starting to set before you added the etbr so uptake was irregular. Try adding it when the gel is much hotter and mix it well before pouring the gel. Incidentally you seem to have a strong primer dimer band in your amplifications which will make it very difficult to get a good amplimer as expected because the primer dimer is removing all of the primer from the reaction mixture
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