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From 2014.igem.org

Electro elution of DNA from agarose gel

Firstly, launch your sample in an agarose gel and migrate. After the migration, cut and save the band containing DNA of interest in an Eppendorf (4°C).

Then, prepare the tank and the salt bridge for the elution :

- Put the specific membrane which will retain DNA in EDTA 500mM (5mL vf) and then put into the microwave for 2mn

- Install the two membranes on the salt bridge and lock them with the rings

- Install the salt bridge between the two tanks

- fill the tank and the salt bridge with TAE 0,1X

- cut the agarose band in little pieces and add them into the bigger receptacle of the saline bridge

- add one drop of BET (0,625mg/mL) in the tank, at the opposite side of your sample

- Connect the generator and migrate at 400V for 30 minutes

After the migration, remove TAE from the saline bridge and check the presence of DNA with a quick UV exposure. If your DNA is at the right side, migrate for few seconds on the other side to detach a little bit the DNA from the membrane. After that, remove the excess of TAE from the saline bridge without enter in the little receptacle where DNA is. Then, take the TAE containing DNA and save it into a tube.

- add TE or water to have 400 uL

- add 200 uL of phenol/chloroforme

- vortex 30' and centrifugate 10

- save the aqueuse phase

- add 1mL ethanol 100% + 140 uL of sodium acetate (3M ini)

Let the DNA precipitate a -20°C (2h to infinity )

- centrifugate 10mn at 4°C

- remove surnatant

- add 1mL ethanol 70%

- centrifugate 10mn at 4°C

- remove surnatant

- put at 56°C , open tube

- add 20 uL H20

- stock at -20°C