FCCT Biochemistry Lab:Agarose gel electrophoresis

iGEM Protocols

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Protocols Making Gel for gel electrophoresis Make 100ml of gel at at time (e.g. 0.8%, 1%, 1.5%, or 2%) Measure out the relevant percentage in agarose (e.g. 2g of agarose for 100ml of 2% gel). Fill it up with 1xTAE buffer. Microwave on low, constantly taking it out every so often to swirl and mix it. Keep microwaving until clear. Make sure it is completely clear Cool to a temperature that is still hot but still can be held in your hand. If the gel gets too cold and starts to harden, start microwaving again until clear. Add 3uL of ethidium bromide for every 100ml of gel after cooled to the correct tempearature. Mix it well by swirling Pour it onto the molds quickly. Wait at least 1/2hr until using it. Gel electrophoresis Select a relevant % agarose gel based on your own experience (general guide: all <1000: 2%, all >2000: 1% guide). Load samples in wells. Load 7uL of marker into a well. Run in the 13x13cm box at 90V or 100V and 400mA for the desired amount of time. View in the gel viewer machine.