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Team:UNIPV-Pavia/Methods Materials/Electrophoresis
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Revision as of 08:30, 24 June 2009
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Electrophoresis
(estimated time: 2 hours)
Materials needed:
- DNA samples
- Ethidium bromide
- Loading buffer 10x Blue Juice, Invitrogen
- TBE (Tris/Borate/EDTA buffer) 5x (final volume 1 liter)
- 54 gr Tris
- 27.5 gr Borate
- 20 ml EDTA 0.5 M (pH 8)
- Marker (1 kb DNA Ladder, Promega)
- Face mask and gloves
- Electrophoresis apparatus
- Transilluminator
- Prepare agarose gel in 1x TBE buffer
- Add ethidium bromide (using gloves and face mask for your safety):
- 1 µl in the small size agarose gel (70 ml)
- 2 µl in the middle size agarose gel (150 ml)
- 4 µl in the big size agarose gel (250 ml)
- Cast the gel, insert the well-forming comb and let it polymerize
- Add the loading buffer to each sample
- Load the samples and 8 µl of marker
- Set to 70-100 volts and electrophorese for the required amount of time
- Use UV-light to look at the bands
- Take a picture of the gel, if needed (not when bands have to be cut!!!)
- NOTE: when not specified, the marker has the following pattern:
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