Agarose gel electrophoresis

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Revision as of 07:28, 28 September 2011

Materials:

DNA template

Agarose

TAE/TBE buffer

6X loading buffer

Procedure:

1. Prepare your gel according to the size of your DNA template. (a 1% gel works fine for most cases) We use 0.7g of agarose in 70ml of TAE buffer

2. Add 2ul of Ethidium Bromide before you pour your gel into the chamber. (WARNING! ETHIDIUM BROMIDE IS EXTREMELY CANCERIGENOUS!)

3. Mix 5ul of DNA with 2ul of loading buffer by pipetting up and down a couple of times. (MAKE SURE YOUR SAMPLE IS AT THE NEGATIVE END OF THE ELECTROPHORESIS CHAMBER!)

4. Load your samples and appropriate marker into your wells.( placing a dark ruler or paper underneath your chamber helps you see the wells better.) . 5. Apply 85 volts to the chamber for 1 hour. ( if you are in a hurry you can apply more voltage but at the cost of picture quality)

6. Check your gel using a transilluminator or other UV emitting device.