Cytomix

Freshly egressed parasites were passed through a polycarbonate filter with a 3-μm pore size (Whatman), then washed, and resuspended in Cytomix (34 (link)) (10 mM KPO₄, 120 mM KCl, 150 mM CaCl₂, 5 mM MgCl₂, 25 mM HEPES, 2 mM EDTA) to 8 × 10⁷ cells/ml. Two hundred forty-five microliters of the parasite suspension was combined with 50 μg of previously linearized plasmid in 155 μl Cytomix, supplemented with 2 mM ATP and 5 mM glutathione (GSH), to a final volume of 400 μl. Parasites were electroporated in 4-mm-gap cuvettes (BTX Harvard Apparatus model no. 640) in an Electro Square Porator (BTX Harvard Apparatus) set to 1.7 kV, for two 176-μs pulses at 100-ms intervals.

Freshly egressed parasites were passed through a polycarbonate filter with a 3-μm pore size (Whatman) and then washed and resuspended in Cytomix (38 (link)) (10 mM KPO₄, 120 mM KCl, 150 mM CaCl₂, 5 mM MgCl₂, 25 mM HEPES, 2 mM EDTA) to 8 × 10⁷ cells/ml. Two hundred forty-five microliters of the parasite suspension was combined with 50 μg of previously linearized plasmid in 155 μl Cytomix, supplemented with 2 mM ATP, 5 mM glutathione (GSH), to a final volume of 400 μl. Parasites were electroporated in 4-mm-gap cuvettes (BTX Harvard Apparatus model no. 640) in an Electro Square Porator (BTX Harvard Apparatus) set to 1.7 kV, for two 176-μs pulses at 100-ms intervals.