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Agarose Mini-Gels 1. In 250 ml flask: Hoefer and Pharmacia BRL

Summary: 28
Agarose Mini-Gels
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 50-55C on bench for 10-15 minutes. DO NOT allow to cool
below 50C. Add 50 l 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 4-25C.
BRL: Do not cool agarose. Add 25 l 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 g/ml EtBr. Submerge gel completely. Remove comb and wash
out wells with buffer using Pasteur pipette.


Source: Aris, John P. - Department of Anatomy and Cell Biology, University of Florida


Collections: Biology and Medicine