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

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We recommend using conditions previously reported to transform a similar biological system as a starting point for the optimization of Biolistic transformation. For example, Armaleo, et al. (1990) report the use of 1.1 mm tungsten particles (equivalent to M17 tungsten) and a vacuum of 0.03 atmospheres (equivalent to 29 inches Hg) to transform yeast cells with the gunpowder-powered PDS-1000 unit (the precursor to the PDS-1000/He system). These parameters were used as starting conditions for optimizing bombardment of yeast with the helium- driven PDS-1000/He system. Preliminary experiments had indicated that a helium pressure of 1,300 pounds per square inch (psi) would produce large numbers of transformants (R. Arentzen, personal communication). Starting conditions for transformation of cauliflower sections were chosen as 1 mm diameter gold or M17 tungsten microparticles and a helium pressure of 1,300 psi, based on experiments using other plant systems (T. Klein, pers. comm.).

Unless otherwise noted, the following parameters were kept constant in all of the experiments described here:

These conditions have proven to be standard for Biolistic experiments involving most cell types (J. Sanford, pers. comm.).

Both gold and tungsten microparticles were prepared, and coated with supercoiled plasmid essentially as described by Sanford, et al. (1992; see Appendices 1 and 2). Microparticles were dried onto macrocarriers in individual desiccators (Petri dishes containing CaSO4) immediately upon application, and were used within 2 hours of drying.


Results
Cauliflower
An example of cauliflower sections transformed with the p
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