Science Buddies: "Ask an Expert"

Hi,

I got prepared 2% agarose and TBE buffer 5X. Will this work following the instructions for the electrophoresis experiment you have on your site or do I need to reorder the exact products you have listed?
I am also unclear as to where to insert the electrodes. When you refer to the top and the bottom of the box do you mean that I put them inside after the gel is poured and solidified at each end of the box?

Thanks for your help!

Hi,

Scibuddies has two electrophoresis projects and I can’t tell from your query which one you are doing:

https://www.sciencebuddies.org/science- ... #procedure
https://www.sciencebuddies.org/science- ... #procedure

I don’t think it really matters, though, as both setups are similar. Agarose gels are usually 1 to 2% agarose, the higher percentage gels being used for separating smaller compounds. Is your stock of 2% agarose in water or buffer? If it is water you will have to melt it in the microwave then add one-fifth volume of 5X TBE to make the buffer concentration 1X. I think TBE is fine to use in place of the specified buffer.

After you pour the gel and it has hardened (about 30 min), cut an 0.5 cm slice from the top and bottom of the gel. This provides a place for the electrodes. Make sure the electrodes are attached securely to the gel box before you start the run and when you attach the batteries, make sure the positive lead is attached to the electrode at the bottom of the gel (the end opposite from the wells).

Be sure to take pictures of your setup and close-up photos of the color bands as they migrate down the gel.

Let us know if you have more questions.

Good luck,

Sybee