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Optimization of Biolistic Transformation Using the Helium-Driven PDS-1000/He System

distance (Table 6). In addition, using smaller microparticles and reducing the chamber vacuum may result in less tissue damage. Initial starting conditions which may be used for the PDS-1000/He with various types of biological systems are given in Table 7. These conditions, though probably not optimal, may be used in any initial experiments to assess optimal biological parameters (e.g., expression vector, cell type, etc.) prior to optimizing the conditions described here.

Finally, the helium pressures reported here are all based on the rating of the rupture disk which was used in each bombardment. The actual pressures at which the rupture disks burst were monitored using the gauge on the PDS-1000/He in approximately half of the 500+ bombardments performed in the course of these experiments. In all cases, the disks burst within 100 psi of their rated pressure. In order to correctly monitor the pressure at which the rupture disks burst, it is important to set the helium pressure at the regulator valve 200 psi over the pressure rating of the rupture disk being used, and to adjust the helium metering valve so that the helium flow rate into the gas acceleration tube occurs at a rate such that a 1,550 psi rupture disk will burst in 12 + 1 seconds. Additionally, in all of the bombardments performed in this study, there was only one rupture disk failure. In order to obtain this low failure rate, it is very important to tighten the rupture disk retaining cap onto the gas acceleration tube as much as possible (the torque on the cap should be between 40 and 50 in-lb).


Conclusions
Stable expression in yeast and transient expression in cauliflower tissue sectio
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