Magnetofection-Based Transfection In Vitro: A Magnetic Field-Assisted Technique to Deliver Plasmids Into Primary Mouse Neuronal Cells Using Magnetic Nanoparticles
Transcrição
To culture motor neurons, dilute them to 5,000 to 10,000 cells in 500 microliters of culture medium per well in a 24-well plate. Then, remove the laminate solution with a P1000 pipette tip. Immediately transfer the motor neurons diluted in culture medium to the coating plates to avoid drying.
To prepare DNA, resuspend 1 microgram of DNA in 50 microliters of neuronal culture medium, and vortex for 5 seconds. To prepare the bead tube, resuspend 1.5 microliters of beads in 50 microliters of neuronal culture medium. Add 50 microliters of bead solution to 50 microliters of DNA solution, and incubate for 20 minutes at room temperature.
During this incubation, withdraw 100 microliters of culture medium from the well that will be transfected. Subsequently, transfer 100 microliters of the DNA-bead mix to each well, and incubate the 24-well plate 20 to 30 minutes on the magnetic plate at 37 degrees Celsius.
