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All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. Induction of Tetracycline-dependent Gene or shRNA Expression
CAUTION: Doxycycline may be harmful. Please refer to the safety data sheet.
NOTE: Depending on the type and duration of the experiment, doxycycline can be supplied in drinking water.
- For short term experiments (<10 days) administer doxycycline via drinking water using the following protocol.
- Dissolve 5 g sucrose in 100 mL of tap water. Autoclave.
- Dissolve 100 mg of doxycycline-hyclate in 5 mL of sucrose solution (step 1.1.1) in a 15-mL conical tube.
- Using a 10-mL syringe, sterile filter the solution through a 0.2 µm filter. Add to the sucrose solution prepared in step 1.1.1.
- Supply doxycycline-sucrose solution as drinking water to the mouse. Check daily and replace when the solution becomes cloudy indicating bacterial overgrowth (replace clear solution after three days at the latest).