BIO RAD Helios Gene Gun System, Инструкция по эксплуатации

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Section 6 Preparation of Mammalian Cell Targets
Helios Gene Gun System | 33
6. Prepare the Helios Gene Gun for operation as described in Particle Delivery Using the Helios Gene Gun,
in Section 5.
Fig. 18. Correct placement of the Gene Gun when transfecting cells in vitro (photo courtesy of Auragen, Inc.).
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.
In Vitro Delivery to Suspension Cultures
Method 1
1. Prepare the Helios Gene Gun for operation as described in Particle Delivery Using the Helios Gene Gun,
in Section 5.
2. Transfer the cells to a centrifuge tube and pellet in a table top centrifuge for 5 min at 250 x g.
3. Resuspend the cells in tissue culture media containing 25 mM HEPES (pH 7.3) at a concentration of
5 x 10
7
cells/ml.
4. Inoculate 20 µl (1 x 10
6
cells) of the cell suspension into a 1.5 cm circle in the center of a 35 mm dish. It
is useful to draw a 1.5 cm diameter circle on an index card for use as a template under the culture dish.