Team:UNIPV-Pavia/Methods Materials/Electrophoresis

(Difference between revisions)

Latest revision as of 10:30, 21 October 2009

Protocols

(estimated time: 2 hours)

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!!!)

INGREDIENTS:

DNA samples

Ethidium bromide

Loading buffer 10x Blue Juice, Invitrogen

TBE (Tris/Borate/EDTA buffer) 5x (final volume 1 liter)

20 ml EDTA 0.5 M (pH 8)

Marker (1 kb DNA Ladder, Promega)

Face mask and gloves

Electrophoresis apparatus

Transilluminator

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 *Take a picture of the gel, if needed (not when bands have to be cut!!!)
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