Team:Paris Saclay/Protocols/Electro elution of DNA from agarose gel


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Electro elution of DNA from agarose gel

  1. 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.
  2. Fill the vat with TAE 0.5X buffer
  3. Close one of the wells of the saline bridge with a semi-permeable membrane, put the DNA in the large well
  4. Apply a 400V current after closing the vat, for 30min
  5. Reverse the polarity for 20s, pipet the solution from the small well and put it in a clean 1.5 ml tube
  6. Add 200µL of water and 600µL of phenol/chloroform/isoamyl alcohol ratio 25/24/1
  7. Vortex for 1 min
  8. Pipet the aqueous phase
  9. Precipitate the DNA with ethanol