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1. Set the electroporation apparatus to 2.5 kV.
2. Add 2 µl plasmid DNA to tubes containing 40 µl fresh or thawed cells on ice. Mix by swirling with pipette tip.
3. Transfer the DNA and cells to a pre-chilled electroporation cuvette (0.2 cm electrode gap) using a narrow pipette tip. Wipe any ice or water from sides of cuvette using a Kimwipe. Place the cuvette into the sample chamber.
4. Energize the electroporation apparatus and deliver the pulse by pushing in both charging buttons simultaneously and holding until a short beep is heard. Note the time constant of the pulse and the actual voltage delivered.
5. Remove the cuvette from the sample chamber. Immediately add 1 ml LB medium and transfer the cells to a sterile polypropylene culture tube using a Pasture pipette. (Failure to immediately add SOC to the electroporated cells can significantly reduce cell viability and decrease transformation efficiency.)
6. Incubate cultures for 60 to 180 minutes at 37°C on a roller or with moderate shaking to allow for plasmid expression.
7. Plate aliquots of the electroporation mixture on L-agar plates supplemented with the appropriate antibiotics. Incubate plates at 37°C.
Protocol adapted from: http://wheat.pw.usda.gov/~lazo/methods/goldberg/electro.html
1. Keep the E. coli on ice for 30 minutes
2. Add 10μl ligation reaction
3. Keep the E. coli on ice for more 30 minutes
4. 42ºC for 1`30``
5. Put in the ice immediately for 5 minutes.
6. Add 500μl LB media
7. 37ºC for 1 hour
8. Plate aliquots of the mixture on LB plates supplemented with the appropriate antibiotics. Incubate plates at 37°C.