BIOLISTIC LAB - Biolistic PDS-1000/He Particle Delivery...

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1 Biolistic ® PDS-1000/He Particle Delivery System Demonstration Introduction Biolistic particle delivery is a method of transformation that uses helium pressure to introduce DNA-coated microcarriers into cells. Microprojectile bombardment can transform such diverse targets as bacterial, fungal, insect, plant, and animal cells and intracellular organelles. Particle delivery is a convenient method for transforming intact cells in culture since minimal pre- or post-bombardment manipulation is necessary. In addition, this technique is much easier and faster to perform than the tedious task of microinjection. Both stable and transient transformations are possible with the Biolistic particle delivery system. The Biolistic PDS-1000/He instrument uses pressurized helium to accelerate sub- cellular sized microprojectiles (gold or tungsten) coated with DNA (or other biological material) over a range of velocities necessary to optimally transform many different cell types. The system consists of the bombardment chamber (main unit), connective tubing for attachment to vacuum source, and all components necessary for attachment and delivery of high pressure helium to the main unit (helium regulator, solenoid valve, and connective tubing). Figure 1. Biolistic components.
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2 Figure 2. Bombardment chamber components Power Switch, ON/OFF Controls supply of line electrical power to the instrument. Vac/Vent/Hold Switch Controls application of vacuum to bombardment chamber. Vac applies vacuum from line source. Vent releases vacuum using filtered air. Hold maintains vacuum by isolating chamber. Bombardments should be performed with this switch in “Hold” position. Fire Switch Controls flow of helium into Gas Acceleration Tube by activating Solenoid Valve. Illuminated red when enabled. Fire Switch must be held ON continuously until Rupture Disk bursts; then release Fire Switch to stop flow of helium. Vacuum Gauge Indicates level of vacuum in bombardment chamber. Vacuum/Vent Rate Control Valves Regulate rate of application and relief of vacuum in bombardment chamber. Clockwise rotation closes valves. Helium Pressure Gauge Indicates helium pressure in Gas Acceleration Tube. Bombardment Chamber Door Closes chamber with a solid piece of polycarbonate plastic. Rupture Disk Retaining Cap Seals Rupture Disk against chamber end of Gas Acceleration Tube. This must be tightened securely. The Torque Wrench is used in the holes in the cap. Microcarrier Launch Assembly Holds the DNA/microcarrier preparation on a Macrocarrier sheet over the Stopping Screen in the path of the helium shock wave. Target Shelf Holds the biological target in a Petri plate in the path of the accelerated DNA/microcarrier preparation. Particle flight distance is determined by positioning the shelf at one of four levels using slots in the chamber walls.
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3 The Biolistic Process The Biolistic PDS-1000/He system uses high-pressure helium, released by a rupture disk, and partial vacuum to propel a macrocarrier sheet loaded with millions of
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This note was uploaded on 07/23/2011 for the course HOS 6737c taught by Professor Moore during the Spring '09 term at University of Florida.

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BIOLISTIC LAB - Biolistic PDS-1000/He Particle Delivery...

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