BIO RAD Helios Gene Gun System Instruction Manual page 38

Section 6 Preparation of Mammalian Cell Targets
5. Hold the dish perpendicular to the spacer and touch the end of the plastic spacer (if the spacer is
sterile), or as close as possible to the target, and discharge the Gene Gun (Figure 18).
6. Add 1.5–2 ml of media to the plate and return to the incubator.
Method 2
1. Prepare 35 mm tissue culture plates with Cell-Tak (Thermo Fisher Scientific, Waltham, MA) using
manufacturer's instructions.
2. Transfer cells to a centrifuge tube and pellet in a table top centrifuge for 5 min at 250 x g.
3. Resuspend cells in tissue culture media without serum at a concentration such that inoculating 2 ml
of cells into the 35 mm tissue culture plates treated with Cell-Tak will produce a monolayer 60–80%
confluent.
4. Inoculate 2 ml of cells into 35 mm tissue culture plates.
5. Incubate 30 min to 4 hr under the appropriate conditions.
6. Prepare the Helios Gene Gun for operation as described in Particle Delivery Using the Helios Gene Gun,
in Section 5.
7. Immediately prior to DNA delivery, aspirate the media from the dish.
8. Hold the dish perpendicular to the spacer and touch the end of the plastic spacer (if the spacer is
sterile), or as close as possible to the target, and discharge the Gene Gun (Figure 18).
9. Add 1.5–2 ml of media to the plate and return to the incubator.
34 | Helios Gene Gun System