Nucleofection of Parasite Forms: An Electroporation-Based Transfection Method to Deliver Fluorescent Protein Encoding-Plasmids Into Sporozoite Nucleus

First, centrifuge the sporozoites or merozoites suspension at 600 x g for 10 minutes, then, discard the supernatant. In the following order, add 85 microliters of nuclear transfection buffer, 10 micrograms of plasmid, and 25 units of restriction enzyme into the 1.5-milliliter tube containing sporozoites or merozoites. Transfer the suspension to a nuclear transfection cup.

Put the cup into a nuclear transfer groove, turn on the nucleofection device by using the power button, and select the transfection procedure U-033. When the program finishes, press the X button of the nucleofection device, and the screen displays OK, indicating that the nucleofection is successful. Add 0.5 to 1 milliliter of Dulbecco's Modified Eagle's Medium to the nucleofection cup to stop the reaction.

After mixing gently, transfer the suspension to a 1.5-milliliter tube.