2. Gel Electrophoresis

3. What is Gel Electrophoresis? Gel electrophoresis separates molecules on the basis of their charge and size. The charged macromolecules migrate across a span of gel because they are placed in an electrical field. The gel acts as a sieve to slow the passage of molecules according to their size and shape. Gel electrophoresis separates molecules on the basis of their charge and size. The charged macromolecules migrate across a span of gel because they are placed in an electrical field. The gel acts as a sieve to slow the passage of molecules according to their size and shape.

4. How does electrophoresis work? The gel is made from agarose. DNA is a negative molecule. Molecules sort based on 1.Charge 2.Size 3.Shape The gel is made from agarose. DNA is a negative molecule. Molecules sort based on 1.Charge 2.Size 3.Shape

5. More Details Because DNA is a negative molecule, when placed in an electrical field it will migrate toward a positive charge, which is the ANODE. +-- Power - +

6. Agarose Gel Used to slow the movement of DNA & to separate it by size Is a polymer extracted from seaweed Sweetened agar gels have been eaten in the Far East since the 17th century. Has no charge! Used to slow the movement of DNA & to separate it by size Is a polymer extracted from seaweed Sweetened agar gels have been eaten in the Far East since the 17th century. Has no charge!

7. Making a Gel An agarose gel is prepared by combing agarose powder and a buffer solution. An agarose gel is prepared by combing agarose powder and a buffer solution. Flask for boiling Buffer Agarose

8. Electrophoresis Equipment Power Supply Gel Tank Casting Tray Gel Combs Cover Electrical leads

9. Preparing the Casting Tray Allow the agarose solution to cool slightly (~60ºC) and then carefully pour the melted agarose solution into the casting tray. Avoid air bubbles.

10. Place the gel in the electrophoresis chamber

11. Add enough electrophoresis buffer to cover the gel to a depth of at least 1 mm. Make sure each well is filled with buffer. Buffer DNA Cathode (negative) Andode (positive)

12. Prepare the Sample Mix DNA samples with loading buffer solution, which contains a tracking dye. Glycerol is also in the solution to help the sample sink into the well. Mix DNA samples with loading buffer solution, which contains a tracking dye. Glycerol is also in the solution to help the sample sink into the well.

13. Loading the Gel

14. Run the Gel Attach power source to gel box. DNA will migrate toward the anode. Attach power source to gel box. DNA will migrate toward the anode. Cathode DNA Anode

15. Analyze the Results

16. Other analysis considerations