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Biology
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Gel Electrophoresis: Lambda Phage

- Understand the process to make a gel.

Jul 28th, 2015

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1. Dissolve the agar, cool the solution, and pour the gel. Combine agar and water. Bring the mixture to a boil and heat until the agar is dissolved. Cool the agar until you can comfortably touch the flask. Place tape across the ends of the gel form (if needed) and place the comb in the form. Pour cooled agar into the form; the bottom 1/3-1/2 of the comb should be immersed. Immediately rinse and fill the agar flask with hot water to dissolve any remaining agar. When the agar has solidified, carefully remove the comb. Remove the tape (if used) from the ends of the gel form.

 2. Load samples in the wells in the gel. Make a written record of which sample you will load in each well of the gel. Place the gel form on a black or dark surface to help you see the wells in the agar. You may find it helpful to only load samples in every other well. Be careful to not puncture the bottoms of the wells 

3. Place the gel in the electrophoresis chamber with the wells closest to the negative (black) electrode. 

4. Prepare the salt solution and add it to the chamber. Add salt to tap water and swirl it to dissolve. Fill each half of the chamber, adding solution until it is close to the top of the gel. Then gently flood the gel from the end opposite the wells to minimize sample diffusion. 

5. Place the lid on the chamber and connect the electrode leads to the power supply. Connect the black lead to the negative terminal and the red lead to the positive terminal. 

6. Turn on the power supply and adjust the voltage to 50-100 volts. 

7. Run the gel for 5-10 minutes. You will be able to observe the samples separating into different colors. Placing the chamber on white paper will help you see the color separation. 

8. Turn off the power supply, disconnect the electrode leads, and remove the chamber lid. 

9. Remove the gel from the electrophoresis chamber. You may also remove the gel from the form and place it on a piece of plastic wrap. Placing the gel on a piece of white paper will help you better see the results. Record and evaluate the results of the electrophoresis. 

10. Clean up. Discard the gel in the trash and pour the salt solution down the drain. Rinse the electrophoresis chamber and gel form with tap water; turn them upside down to dry

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Jul 28th, 2015

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