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Talk:Team:EPF-Lausanne/Protocols
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Agarose gel electrophoresis
made on 01.07.2011
Purpose
To analyse DNA composition of a sample. Voltage-dependent DNA migration through agarose gel allows sequences separation according to their size.
Ingredients
for a 1% agarose gel
| Small | Medium | Big 26 wells | |
| Agarose | 0.5g | 50ml | 1.25µl |
| TBE 1x buffer | 1g | 100ml | 2.5µl |
| Gel Red | 2g | 200ml | 5µl |
Method
• Weight agarose, place it into a bottle with a cap, add TBE 1x buffer
• Place the bottle into a microwave, <gras>loosen the cap</gras>, heat it untill it almost boils and the agar is dissolved
• Cool it down until it can be hold (that can be done under water)
• Add 1.25µl of Gel Red
• Pour into the rack, put the comb and wait while it polymerizes
Usually “B” valves are starting to load earlier (leave the button unpressurised prior to the pressure increase, it tends to stick to the slide.)
Increase the pressure in both “A” and “B” valves till ~13-15 psi and make sure that you can see that all chambers are separated. If no – increase the pressure
