Gel electrophoresis

 

Prepare Gel

80ml 0.5xTBE + 1.2g Agarose in small glass beaker covered with Al-foil

1. boil, prevent bubbles
2. cool down to ~60oC
3. add 10µl Ethidium Bromide (10µg/µl), stir gently with pipette
4. pour into taped tray, insert combs, remove/move bubbles with pipette
5. wait ~30min

Prepare Samples

Spin down (condensation/sample in lid)

Add 10% (2.5µl to 25µl sample) loading buffer to samples
Vortex + spin down
(DNA size ladder- 1.5µl aliquoted DNA ladder + 1µl loading buffer + 7.5µl ddH2O)

Load/Run Gel

Remove tape on tray

Place Gel/Tray in machine (note wells towards –pool)
Load 10µl/lane
Cover with Cling film, Al-foil + Cardboard box (to prevent evaporation + photbleaching)
Turn on power supply
Capture Image

Place gel on plastic-foil

UV-illuminator, high intensity, green filter
Capture image using Optimas, fixed setting (Brightness-121, Contrast-255)
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