Principles of particle bombardment

The sections below set out important parameters for transformation by particle bombardment and discuss the different bombardment devices available. As 6r as apparatus and procedural matters are concerned, there are no special considerations fbr the delivery and expression of antibody transgenes as compared to transgenes encoding any other type of protein. 

Birch and Bower identified and discussed a number of important parameters for bombardment-mediated gene transfer. A number of these parameters were translated into principles that should be generally applicable in many diverse 

Particle bombardment devices must be safe to operate. 

Particle bombardment-mediated transformation should be of consistent efficiency. 

The ability to direct particles to specific cell groups is an advantage. 

Bower and Birch also formulated a general strategy for optimizing a stable transformation method based on the above principles. The steps they recommended are listed below  

The most important considerations for vector design therefore reflect the structure of the expression cassette. Such considerations (as discussed above) include  

Prepare high quality plasmid DNA at a concentration of 10 mg/ml. We find the Q,iagen maxiprep kit is satisfactory. Concentrate DNA by ethanol precipitation and check concentration by UV spectrophotometry. 

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