2) Heat inactivate ligation mix 15 minutes at 68°C

3) Put 20µl of cells + 1µl of ligation mix into sterile tube

4) Fill electroporation apparatus with ice, put electroporation chamber into slots

5) Suspend cells + DNA between dimples on chamber

6) Charge to 420 volts, arm, then Discharge when it reaches 400 Volts

7) Allow cells to rest about 2 minutes in chambers

8) Add cells to 1ml SOC, 37°C, no antibiotics

9) Shake in 37° incubator 1 hour

10) Plate onto 3 100mm LB-agar plates with appropriate

antibiotics. Plate 1: 100µl of cells

Plate 2: 250µl of cells

Plate 3: Rest of cells

11) Incubate overnight 37° incubator then store @ 4°C no more

than 4 weeks before re-plating

Settings on BRL Electroporator Model 1600

Power Supply: Capacitance......... 330µF

DC Volts................ Low ½

Charge Rate......... Fast

Voltage Booster k½................ 4