Electroporation Protocol

  1. Obtain a competent cell aliquot from the -80oC freezer, keep the tubes on ice.
  2. Add 40μL of ice cold H2O to each aliquot to make a 80 μL solution.
  3. Separate the 80 uL solution into 2 separate tubes to make two 40uL tubes of solution.
  4. Add 1μL of a Gibson Product into each tube
  5. Using the Electroporator:
    • Set the electroporator to 1250 V. and press "time constant".
    • Obtain a chilled cuvette, and pipet all 40 uL of a sample into the center.
    • Place the cuvette into the electroporator and press "Pluse" twice.
    • Immediately remove the cuvette and rescue the sample in 300-500 mL of LB.
    • Pipet the sample into a labeled microcentrifuge tube, and record the time constant. (The time constant should have a value greater than 2.5).
      • Repeat this process for the other sample. After the addition of LB, transfer the sample into another labeled microcentrifuge tube.
  6. Incubate both samples for ~1 hour @ 37oC.
    • After incubation, combine both samples (make sure each sample has a comparable time constant) and follow standard plating procedures.