Nucleofector PF Protocol

  1. Collect 4x10e7 procyclic form cells, spin down 1300xg, 10 min.
  2. Resuspend cells in 100μl room temp parasite kit 1 Nucleofector solution (that had the supplement added).
  3. Mix this solution with 10μg DNA.
  4. Transfer cells/DNA into certified cuvette (make sure there is no air bubbles).
  5. Select nucleofector program X-0001.
  6. Insert the cuvette into the nucleofector cuvette holder and apply selected program.
  7. Take the cuvette out of the holder, add 500μl of equilibrated culture medium (w/15% FBS) and transfer the sample into a culture flask.
  8. Add to 30ml media.
  9. Make 1:20 dilution (1.5ml +28.5ml media).
  10. Aliquot 1ml of each into 24 well plate.
  11. Let sit in incubator overnight and add 1ml of media +2X selection into each well.

Download Protocol: Nucleofector PF Protocol
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