Collect 4x107 procyclic form cells, spin down 1300xg, 10’.
Resuspend cells in 100ul room temp parasite kit 1 Nucleofector solution (that had the supplement
added).
Mix this solution with 10ug DNA.
Transfer cells/DNA into certified cuvette (make sure there is no air bubbles).
Select nucleofector program X-0001.
Insert the cuvette into the nucleofector cuvette holder and apply selected program.
Take the cuvette out of the holder, add 500ul of equilibrated culture medium (w/15% FBS) and transfer
the sample into a culture flask.
Add to 30ml media.
Make 1:20 dilution (1.5ml +28.5ml media).
Aliquot 1ml of each into 24 well plate.
Let sit in incubator overnight and add 1ml of media +2X selection into each well.