Author: Garrick Wilson
Magnet Assisted Transfection of Adherent cells (MATra-A)
Protocol is optimised for transfection of HepG2 cells seeded into 24 well plates.
1. Seed cells to desired density overnight (recommend 2 x 105 cells/cm2 )
2. For 1 well of a 24 well plate add 600ng of DNA to 50ul of serum free DMEM
3. Add 0.6ul of MATra reagent to the DNA:DMEM mixture, mix thoroughly and
incubate at room temperature for 45mins.
4. During incubation, replace media on adherent cells with 0.5ml DMEM supplemented
with 3% FBS (P/S free).
5. Add the DNA:bead mixture from step 3 to the cells, mix gently
6. Place the plate onto the magnet
7. Incubate for 2-3 hours at 37˚C
8. Perform a media change – particularly if the transfection has been carried out in
serum free media (3% DMEM).
9. Continue to culture cells as desired until evaluation of transfection efficiency (48 -72
hours later)
Important – keep MATra reagent as far away from the magnet as possible,
furthermore keep the reagent away from any strong magnetic forces as this may
agglutinate the nano-particles leading to impaired functioning.
May need to optimise protocol for other cell lines.