1. In 250 ml flask: Hoefer and Pharmacia BRL
50 ml ddH2O 25 ml ddH2O
1 ml 50X TAE buffer 0.5 ml 50X TAE
0.4 g Agarose 0.2 g Agarose
0.8% agarose is good for most purposes.
1.0% agarose is better for resolving DNA bands < 1kb.
2. Place glass cap on flask. Zap in microwave oven at 100% until all agarose is dissolved. Swirl
flask intermittantly. To avoid excess water loss, weigh flask on top-loading balance and add back
lost ddH2O after agarose is dissolved.
3. Hoefer and Pharmacia: Cool agarose to 50-55°C on bench for 10-15 minutes. DO NOT allow to cool
below 50°C. Add 50 µl of 1000X ethidium bromide. Swirl to mix. Avoid creating bubbles. Pour
agarose solution into clean casting stand on level surface. Use clean comb and check distance under
wells (>1 mm). Pop bubbles with Pasteur pipette. Cool for 15-30 minutes. To store gel, cover
with plastic wrap and store at 4-25°C.
BRL: Do not cool agarose. Add 25 µl of 1000X EtBr. Swirl to mix. Pour gel immediately.
· 1000X ethidium bromide stock = 0.5 mg/ml EtBr
4. Set up gel with 1X TAE buffer + 0.5 µg/ml EtBr. Submerge gel completely. Remove comb and wash
out wells with buffer using Pasteur pipette.