Agarose Gel Electrophoresis

Reagents and Materials

1X TA

Graduated Cylinder

125 mL flask

Agarose

Gel Pouring Tray

Tape

Gel rig

Red Safe

Protocol

Measure out 100mL of 1x TAE buffer;

Transfer buffer to 125 mL flask;

Weigh out enough agarose to make a 1% gel (in our case 1.0 g of agarose was the right amount);

Transfer agarose to 125mL flask;

Melt agarose in microwave until solution is almost boiling, stirring every 15-20 seconds (should be around 2 minutes);

Allow agarose to cool (do not let it cool to the point where it is hard);

Add 5 uL of Red Safe to the cooling agarose;

Assemble the gel pouring apparatus by inserting gate into slots;

Allow gel to cool until flask can be handled comfortably;

Place comb in the gel rig;

Pour agarose into gel tray;

Allow to solidify. While the gel is solidifying prepare the samples. Add your sample and 1 uL 10x Loading Dye, 4 uL of DNA and 5 uL of water;

Pour 1X TAE over gel so that gel is covered by a 3-5mm buffer;

Load samples into lane (Don't forget to load a 1kb+ ladder into one of the lanes);

Hook electrodes to gel apparatus;

Run the apparatus at 100V for 30 - 45 minutes (make sure to watch that the dye does not run off the gel);

Visualize the gel and record the results.