Procedure

1. Equipment Required:

• gene gun (helium-driven)
• Microcarrier particles (gold or tungsten)
• Genetic material (DNA or RNA)
• A breach disk
• Macrocarrier and stopping screen
• Target cells (plant or animal cells)
• A Petri dish or growth plate
• A micropipette
• A vacuum chamber (optional)
• Pressurized helium supply

2. Sample Collection:

• Target cells are prepared and placed on a sterile Petri dish or growth plate suitable for bombardment. The cells are ensured to be in optimal growth conditions before proceeding.

3. Sample Preparation:

• The genetic material for transfer is selected (e.g., plasmids, DNA fragments).
• Microcarrier particles are coated with the genetic material using a binding solution (e.g., calcium chloride), and the particles are allowed to dry.
• Uniform coating is ensured to optimize genetic transformation.

4. Mounting the Sample:

• The target cells are securely placed on a Petri dish or growth plate and positioned inside the gene gun’s chamber.
• The surface area of the cells is evenly exposed to ensure efficient bombardment.

5. Equipment Setup:

• The coated microcarrier particles are loaded into the gene gun.
• Pressure settings on the helium tank are adjusted according to the specifications for the target cell type.
• The breach disk is inserted, and the stopping screen is prepared, ensuring proper alignment to stop the macrocarrier while allowing the microcarriers to pass through.
• The vacuum chamber is activated (if used) to prevent air resistance during the bombardment process.

6. Observation:

• Once the bombardment is completed, the cells are transferred from the Petri dish or growth plate to a suitable growth medium.
• The cells are allowed to recover, and signs of genetic transformation are monitored, such as the expression of a reporter gene or changes in cell morphology.
• The cells are observed under a microscope to verify the successful penetration of the microcarriers and delivery of the genetic material into the cytoplasm.