Procedure
1. Chill the electroporation cuvettes by floating them in an ice bath.
2. Remove vials containing 100μl electro-competent cells from the -80°C freezer and thaw them with the iced cuvettes.
3. Prepare micro-centrifuge tubes containing 900μl SOB media.
4. Turn on electroporator and set voltage to 1.5 kV (1mm cuvettes).
5. Add 5μL of ligated DNA sample to 100μl thawed electrocompetent cells on ice. Swirl the tip around gently in cells to mix DNA and cells.
6. Place cells back on ice to ensure they remain cold.
7. Pippette 100μL of cell-DNA mixture to the cuvette.
8. Wipe off excess moisture from outside of the cuvette.
9. Place cuvette in chamber of electroporator.
10. Pulse the cells by pressing the button on the electroporator twice.
11. Quickly use a pipette to remove the electroporated cell suspension from the cuvette and add it to a tube containing 1ml SOB.
12. Let cells recover at room temperature for 30-60 minutes.
13. Plate 100μl of electroporated cells onto a prewarmed LB-agar plate supplemented with appropriate antibiotics. Incubate plate overnight at 37°C.