How to Make an Agarose Gel

1. Weigh out 1 gm of agarose powder. Put the powder into a 250 ml Erlenmeyer flask
2. Add 100 ml of TBE( Tris Borate EDTA) Buffer of TAE ( Tris Acetate) Buffer for every 1 gm of agarose. TBE and TAE are both used for proteins and DNA analysis. In the hemoglobin experiment, TAE buffer was used. In the DNA experiment TBE buffer was used. The pH of TAE buffer is 9.2. The pH of TBE buffer is 8.6. The more alkaline pH is better for the separation of hemoglobin molecules in the gel.
3. Swirl the mixture of powder and buffer.
4. Place in the microwave for 1.5 minutes. Remove with the hot holders and swirl again. Put the mixture into the microwave for another minute. If mixture isn’t clear and bubbling. Try another 1/2minute in the microwave.
5. Remove the flask from the microwave with the hot holders
6. Place on desk top. Allow it to cool to touch.
7. Pour into gel molds. Gel molds should be prepared ahead of time. Place the combs into the gel mold.
8. Allow to cool and solidify.
9. Remove comb and gel. Place the gel in gel box and cover with buffer. If storing the gels place them in a zip lock bag, add several mls. of buffer, and refrigerate until ready to use.

DONT'S

Do not remove combs until the gel is totally solidified. This may affect the wells.

Do not pour hot agar into the gel molds. This ruins the teeth of the comb and can affect the shape of the wells.

Do not pour agar into the gel mnlds if it is beginning to solidify. This creates an uneven gel.