Electroporation

• Dialysis of ligation product first: float a nitrocellulose membrane filter on water in a petri dish with the shiny side up, drop of your 5 µl ligation product on top for 30 min.
• Add 1 µl of ligation product or plasmid (~100 ng/µl) to cells. Place in electroporation cuvette.
• Electroporate (1900 V for 1 mm gap cuvette, 2500 V for 2 mm gap cuvette) and time constant τ = 5 μs.
• Add 900 μl SOC as soon as possible. Be gentle, but mix and place in 1.5 ml Eppendorf tube.
• Shake at 37 °C for 1 hour.
• Plate 200 µl, or alternatively plate all by spinning down and resuspending in ~200 µl of dregs.