Electroporation-Mediated In Vitro Gene Delivery: An Electric Pulse Technique to Introduce Fluorescent Reporter Protein-Encoding Plasmids Into Cultured Cells

To begin, add 0.5 milliliters of culture medium into each wall of the 24-well culture plate containing coated coverslips, and keep it warm at 37 degrees Celsius in a carbon dioxide incubator. Pipette the required number of cells into a sterile 1.5-milliliter microcentrifuge tube and spin at 200 x g for 5 minutes at room temperature.

Discard the supernatant, and resuspend the cell pellet in 200 microliters of Opti-MEM. Repeat this procedure twice to ensure no residual culture medium is present in the tube. Set the parameters of electroporation as listed. Pipette the electroporation reaction gently to mix well, and use a long P200 pipette tip to transfer an exact volume of 100 microliters of the mixture into the two-millimeter gap cuvette.

Once the cuvette is inside the cuvette chamber, press the omega button of the electroporator, and note the impedance value by adhering to a precise volume of 100 microliters. The range of impedance values should be approximately 30 and 35.

Press the Start button to initiate the pulse. Record the measured current values and jewels shown on the reading frame. Remove the cuvette from the chamber.

Immediately add 100 microliters of pre-warmed culture medium into the cuvette, and resuspend it by pipetting up and down two to three times. Immediately transfer the cell suspension to the 24-well plate previously prepared. Incubate the cells at 37 degrees Celsius in a carbon dioxide incubator.