Team:Paris Saclay/Protocols/Electro elution of DNA from agarose gel
From 2014.igem.org
Electro elution of DNA from agarose gel
- Run samples on agarose gel as usual. When fragments are well separated, slice out bands of interest with a razor blade under the UV plate. Make slices as thin as possible (trim off excess agarose).Slices can be stored in microcentrifuge tubes in the refrigerator if necessary.
- Fill the vat with TAE 0.5X buffer
- Close one of the wells of the saline bridge with a semi-permeable membrane, put the DNA in the large well
- Apply a 400V current after closing the vat, for 30min
- Reverse the polarity for 20s, pipet the solution from the small well and put it in a clean 1.5 ml tube
- Add 200µL of water and 600µL of phenol/chloroform/isoamyl alcohol ratio 25/24/1
- Vortex for 1 min
- Pipet the aqueous phase
- Precipitate the DNA with ethanol