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Team:UNIPV-Pavia/Methods Materials/Electrophoresis
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| - | + | =='''Electrophoresis'''== | |
| + | ''(estimated time: 2 hours)'' | ||
| + | <br> | ||
| + | <br> | ||
| + | '''Materials needed:''' | ||
| + | *'''DNA samples''' | ||
| + | *'''Ethidium bromide''' | ||
| + | *'''Loading buffer 10x Blue Juice, Invitrogen''' | ||
| + | *'''TBE (Tris/Borate/EDTA buffer)''' | ||
| + | ** '''TBE 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''' | ||
| + | <br> | ||
| + | *Prepare 1% 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 (220 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 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) | ||
| + | <br> | ||
Revision as of 23:03, 23 June 2009
|
Electrophoresis
(estimated time: 2 hours)
Materials needed:
- DNA samples
- Ethidium bromide
- Loading buffer 10x Blue Juice, Invitrogen
- TBE (Tris/Borate/EDTA buffer)
- TBE 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 1% 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 (220 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 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)
