Encyclopedia of Experiments: Immunology
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Transcript
After anesthetizing the mouse, apply veterinary ophthalmological ointment to its eyes to prevent dryness. Then, shave the skin on the back of the animal and apply a depilatory cream to dissolve keratin proteins in the remaining hair shafts. Following depilation, wipe off the cream with water-soaked cotton.
Using a sterile cotton swab soaked in 70% ethanol, disinfect the skin's surface and mark the target skin area with a previously prepared template. Apply 10 microliters of the DNA solution containing 10 micrograms of plasmid DNA to the desired skin area. To acufect the mouse, hold the acupuncture apparatus perpendicularly to the skin, and repetitively puncture the designated surface either 100 times over 30 seconds or until the solution applied to the skin absorbs.
To achieve successful plasmid DNA expression in the skin tissue, it is important to avoid cutting deep into the skin and to make sure the DNA solution is well-absorbed.
After completing the procedure, place the acufected mouse on a heating pad to maintain its body temperature until recovery from anesthesia. When the mouse regains sufficient consciousness, return it to its cage and provide water and food. Closely monitor the mouse for any symptoms of discomfort, including behavioral changes, abnormal appearance for the first 48 hours after the procedure.
