I would like to know that can we reuse agarose gel containing EtBr by remelting it again since it got solid before it is poured into the gel tray? Do I have to add EtBr again in that agarose gel?
Thank you very much.
Hi Prasansah, it is ok to do it. Just remember to melt it down under a fume hood as etbr fumes are dangerous. You could avoid etbr by switching to methyl green (I know some Indian colleagues are already using it), see https://www.researchgate.net/publication/261139124_A_fast_low_cost_and_highly_efficient_fluorescent_DNA_labeling_method_using_methyl_green .
Keep in mind that the amount of DNA in your original gel will translate into background on the second (remelt) gel, so try to figure out some methods to keep it low (something like running it longer to let DNA out before remelting, etc). A booster of your dye would be recommended.
Please keep us updated!
Best,
D
Hi Prasansah, it is ok to do it. Just remember to melt it down under a fume hood as etbr fumes are dangerous. You could avoid etbr by switching to methyl green (I know some Indian colleagues are already using it), see https://www.researchgate.net/publication/261139124_A_fast_low_cost_and_highly_efficient_fluorescent_DNA_labeling_method_using_methyl_green .
Keep in mind that the amount of DNA in your original gel will translate into background on the second (remelt) gel, so try to figure out some methods to keep it low (something like running it longer to let DNA out before remelting, etc). A booster of your dye would be recommended.
Please keep us updated!
Best,
D
I agree with Daniel except that EtBr fumes are an urban legend. EtBr is not volatile (it is a quaternary ammonium base). However, it is quite correct that boiling EtBr should be performed in a fume hood because it generates EtBr-containing aerosols
I've worked in several labs that would melt and reuse agarose with EtBr. I've even been in a lab where they would stuff a flask with old gels and just scoop out a chunk when they need to pour a gel. You'll have to add in more pure water to make up for volume lost while boiling and add in fresh EtBr every time. Re-used agarose gels tend to have a bit of a background color from the loading dye, existing DNA, existing EtBr, so I always use fresh agarose if I am looking for a small/faint produce or for cloning.
Agree with all the above comments. We reused our gels with EtBr for many years and of course you have to add EtBr every time you melt it (inside the fume hood). Then we switched to red safe instead of EtBr but we still reuse our gels 2-3 times.
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