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Sub-Micron Gold Particles Are Superior to Larger Particles for Efficient Biolistic Transformation of Organelles and Some Cells

) and certain cell types (Chlamydomonas, yeast, and maize callus) occurs using sub-micron sized gold particles. Smaller particles are probably more efficient than larger particles for Biolistic transformation in these systems because they cause less cell damage, resulting in better survival. Organelles could be irreversibly destroyed upon impact with large particles. In the case of maize callus, while cells bombarded with both 0.6 and 1.0 gold particles transiently express equivalent levels of anthocyanin four days postbombardment, cells bombarded with the smaller particles gave rise to many more stable transformants, most likely because they sustained less cell damage which could be more easily repaired.

Advantages of gold microparticles include their uniform size and shape and their biological inertness. In contrast, tungsten microparticles are irregular in shape and heterogeneous in size, can be toxic to cells, are subject to surface oxidation that can alter DNA binding, and, over time, can catalytically degrade DNA bound to them (Sanford, et al., 1993). Thus, because of higher transformation efficiency of gold particles, greater variability in results using tungsten, and the need for special precautions in particle preparation, handling, and transformation procedures when using tungsten, gold particles are superior to tungsten particles for Biolistic applications. 0.6 gold microparticles can be used with the Yeast Optimization Kit (catalog number 170-3100) to introduce the novice to the principles of particle bombardment.

The size of small particles may be determined by several methods, including molecular sieving, sedimentation, light scattering, electrical sensing, optical sensing, and
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