Agarose Gel 0.

1. Fill a flask with 100mL of 1X TBE buffer 2. Using a weigh boat, first zero out the weigh amount by tearing the scale. This is done by placing the weigh boat on the scale and pressing 0/T button until the reading displays 0. 3. Weigh 0.7g of agarose (powder form) and pour into flask containing the 1XTBE buffer 4. Place a kimwipe just inside the top of the beaker to avoid any boiling over and place in microwave 5. Heat solution for approximately 1.5 minutes, swirl beaker and heat again for approximately 30 seconds or until the solution boils. The agarose will not solidify unless it reaches the boiling point. 6. Let the solution cool to the touch and add 10.0µL of EtBr into the solution. **it’s important to allow solution to cool, you do not want to breathe the vapors of EtBr. Swirl beaker 7. Place gel tray into gel rig with the gaskets sealing in the tray tight so that no agarose escapes. See figure A 8. Place gel comb into provided slot 9. Pour agarose into gel tray slowly to avoid bubbles. Allow to solidify. 10. When gel is solid, take out the gel tray, with the solidified gel, and turn it to fit into gel rig with the comb end on the side of the black (or negative) end of the rig. 11. Fill a graduated cylinder with 1X TBE buffer and pour it into the gel rig filling it to the fill line ensuring that the gel is covered with the buffer. 12. Pull out the comb, slowly and straight up to avoid tearing of the gel. 13. To your reaction, add 1.0 µL of 1X loading dye and pipette up and down to mix 14. Make a list or chart of the gel and mark which well you added what reaction tube Ex: lane 1 – DNA ladder Lane 2 – reaction 2 15. Add 3 µL of DNA ladder to lane #1 on your gel 16. For each well, add 10 µL of your reaction with the dye included 17. After gel is loaded, place the top of the rig onto the box making sure that the colors match for the electrodes and seal tightly 18. Set voltage to _________ and time to _________

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