Gel Electrophoresis

Gel Electrophoresis

(A) Agar Gel preparation
1. Put 0.9 g agarose in a 250ml flask with 70ml 1X TBE(a buffer solution)
2. boil it until it becomes clear
3. allow the gel solution to cool to about 50 oC

(B) Setting the running chamber
4. Use the plastic paper to make the tank become sealed. Set the comb over the gel plate so that the teeth rest just above the plate.
5. slowly pour the gel onto the plate until the solution covers about one-half the height of the comb’s teeth. (avoiding creating any bubbles)
6. allow the gel to solidify.
7. when the gel has hardened, squirt some water around the comb and then pull the comb teeth from the gel slowly.

(C) Loading the samples
8. place your gel in the running chamber with wells nearest the negative(black) electrode, and completely cover the gel with 1X TBE solution. DO NOT move the running chamber from this point on.
9. carefully load each of the samples(MK, M, F, S1, S2, D1, D2) into their corresponding wells in your gel.
10. attach the positive(red) and negative(black) electrodes to the corresponding terminals.(red into red, black into black) on the power supply and on the gel box.
11. turn the power on to about 100-120 volts, and make sure that small bubbles arise from the electrodes in the gel buffer.
12. wait for about one hour or the samples has moved about 3/4 distance .

(D) Staining the samples
13. Put the gel out of the tank and put it into the staining solution overnight.