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Team:Tuebingen/Notebook/Protocols/gelelectrophoresis
From 2013.igem.org
Reagents
| 1.0 g | Agarose |
| 100 mL | 1x TAE (dilute 50x TAE accordingly) |
| 1.7 µL | GelRed |
| 1.0 µL | 1x Loading buffer (dilute with Aqua dest. if necessary) per well |
| 10.0 µL | Ladder-buffer mix per well |
| 5.0 µL | DNA sample per well |
Procedure
Add agarose to TAE buffer and cook in microwave until completely dissolved. Add GelRed to the liquid (cooling down is not necessary). Prepare casting of gel and let liquid cool down a bit. Cast gel and let solidify. Place solid gel in gel electrophoresis apparatus (wells should be near cathode because DNA moves towards anode) and fill with 1x TAE buffer (see manual of gel electrophoresis apparatus for ideal / maximum amount of buffer).
Fill desired amount of wells with 10 µL ladder-buffer mix each. For each well mix 5 µL DNA sample with 1 µL 1x loading buffer and fill 5 µL of this mixture in well.
Start gelelectrophoresis by applying 80 V DC. After 10 min increase voltage to 100 V. Stop gelelectrophoresis when marker reaches bottom of gel.
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