DNA Gel Electrophoresis
An.Huang
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Abstract
Agarose gel electrophoresis can help determine the mass of a certain DNA fragment. This protocol will help user conduct gel electrophoresis appropriately.
Steps
Mix 1g agarose with 100mL electrophoresis buffer (1X TAE) in a flask, then heat and stir until agarose is completely dissolved.
Wait until the molten agarose cools down to around 40°C .
While waiting for the cool down of molten agarose, set up and level the gel casting tray and the gel casting platform on a flat area. Inset the comb.
Add 10µL 10000X GelRed nucleic acid stain into molten agarose and mix well.
Pour the molten agarose onto the tray. Immediately rinse out the flask before any remaining agarose sets in it.
Wait at least 0h 20m 0s for gel solidification.
While waiting for gel solidification, prepare the samples as follows:
Mixing the components:
6X gel loading dye X μL
DNA sample/DNA ladder 5X μL
Once the gel has completely solidified, gently remove the comb by pulling it up slowly and vertically.
Carefully place the tray in the gel tank. Add electrophoresis buffer (1X TAE) until covering the whole gel surface.
Pipette the prepared samples using a P20 (2μL-20μL) pipette to the wells.
Run the gels at 90 V, approximately 50 mA, for 90 min.
When the run is finished, carefully take the gel tray with gel out from gel tank. Photograph the gel under ultraviolet (UV) light.
