Part C

Running a Gel

13. Retrieve your microtubes (A-E) and your gel. If the materials in the microtubes are frozen, hold each tube in your hand until the solutions thaw.

14. Set a micropipetter to 1 pL, and place a tip on the end. Add 1pL of loading dye to each microtube. Use a new tip for each microtube. Gently tap each microtube on your lab table to thoroughly mix the solutions. Do not shake the microtubes.

15. Remove your gel (still in the gel-casting tray) from the plastic bag, and place it in a gel chamber. Orient the gel so that the wells are closest to the black wire, or anode.

16. Set a micropipetter to 10 pL, and place a new tip on the end. Open microtube A, and remove 10 pL of solution. Carefully place the solution into the well in lane 1, the left-most lane. To do this, place both elbows on the lab table, lean over the gel, and slowly lower the micropipetter tip into the opening of the well before depressing the plunger. Do not jab the micropipetter tip through the bottom of the well.

17. Using a new micropipetter tip for each tube, repeat step 16 for each of the remaining microtubes. Use lane 2 for microtube B, lane 3 for microtube C, lane 4 for microtube D, and lane 5 for microtube E.

18. Very slowly fill the gel chamber with 1 x TBE buffer until the level of the buffer is approximately 1-2 mm above the surface of the gel.

19. CAUTION Follow all of the manufacturer's precautions regarding the use of this equipment. Close the gel chamber, and connect it to a power supply according to your teacher's instructions.

20. Allow an electric current to flow through the gel. You will see a blue line moving away from the wells. When the blue line is approximately 5 mm from the end of the gel, disconnect the power supply and remove the gel. Store the gel overnight in the plastic bag.

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