Gel Electrophoresis Overview
Gel Prep
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Grab 250 mL flask; measure and pour 50 mL of 1x TAE Buffer into it
Measure out 0.5 grams of agarose and pour into 50 mL of TAE
Be sure to tape up gel casting trays and have an 8-well comb placed inside
Heat up gel mixture (agarose w/TAE) in microwave
a. Solution should lose all precipitate when properly heated
b. Bring heated flask to sink and cool under water
5. Once gel solution is cooled enough to pour, add 4 uL Ethidium bromide to solution and swirl to
properly mix EtBr
6. Pour gel mixture into casting tray and allow to cool *be sure to move out air bubbles*
DNA Prep
1. While gel is cooling prepare DNA samples for running
2. Each sample of DNA will contain 10 uL of DNA from PCR amplification
a. Samples will be prepared for you, be sure to mark down sample name and tube number
3. Add too each sample 1.1 uL of 10x DNA Loading Dye
a. Mix loading dye directly into sample and pipette up and down (example demonstration
if needed)
b. Use pipette to mix sample and dye, do not flick tube or shake tube
4. Do this for each sample
Load DNA Gel
1. Place solidified gel into electrophoresis tray and fill with 1x TAE ( be sure buffer is higher than
top of gel; but not too high that leakage occurs from the top)
2. Add 5 uL DNA ladder to your gel
3. Add each sample to each well
4. Add 3-4 uL EtBR into TAE buffer in tray (this step will be discussed in greater detail during lab)
5. Run gel at 100 Volts until yellow dye band is close to ¾ of the way down the gel
6. Stop run at this point