SBI4U1
May 13th, 2013
Gel Electrophoresis Lab
Purpose:
The purpose of this lab is to learn how restriction enzymes cut DNA molecules at specific sequences, thus producing DNA fragments of various lengths. Students learn how fragments form unique patterns, which help to distinguish the base for DNA identification. This lab answers the question “whose DNA was left behind?”.
Materials: * Transfer pipets
* Agarose Gel
* Dyed DNA samples
* Electrophoresis Buffer
* Electrophoresis Chamber
* Casting Tray
* Dams
* Comb
* Power Supply
* Heat Source
* 500 ml Beaker or Flask
* Distilled Water (used to make buffer solutions)
Procedure:
1. After all of the required materials are received, rubber dams must close off the ends of the casting tray.
2. A comb is placed in the first set of notches nearest the end of the casting tray, making sure it sits evenly in the tray.
3. The previously prepared agrose gel is then poured into one end of the casting tray on a level surface until an even layer is spread across the bottom of the tray.
4. After the gel has solidified, the dams must be carefully removed from the gel.
5. After the dams are removed, slowly remove the comb from the gel by pulling straight up.
6. Place the tray in center of the chamber, and fill the chamber with the required amount of distilled buffer solution.
7. Holes must be poked into the dyed DNA samples using a pencil.
8. Using the pipette, begin to take a sample of “DNA A” whilst squeezing the pipette approximately 1/3 of the way to the bulb.
9. Insert “DNA A” into lane 1, and repeat this process until B-2, C-3, D-4, E-5, F-6.
10. After samples are loaded, place the cover on the chamber making sure the positive and negative ends are correctly oriented.
11. Insert the plugs and wire with the coordinating colours and attach to the power source.
12. Set the power