Nucleofector PF Protocol
- Collect 4x10e7 procyclic form cells, spin down 1300xg, 10 min.
- Resuspend cells in 100μl room temp parasite kit 1 Nucleofector solution (that had the supplement added).
- Mix this solution with 10μg 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 500μl 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.
Download Protocol: Nucleofector PF Protocol
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