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JoVE Encyclopedia of Experiments
Encyclopedia of Experiments: Biological Techniques

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Inducible Tet-On Regulatable System-Based Gene Expression In Vivo

 

Inducible Tet-On Regulatable System-Based Gene Expression In Vivo: A Tetracycline-Induced Gene Expression System for Target Gene Transcription Activation in Mouse Model

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For experiments of less than 10 days, continuously administer doxycycline via drinking water using the following protocol. First, dissolve 5 grams of sucrose in 100 milliliters of tap water, and autoclave.

Then, dissolve 100 milligrams of doxycycline-hyclate in 5 milliliters of the sucrose solution in a 15-milliliter conical tube. Next, use a 10-milliliter syringe to sterile filter the solution through a 0.2-micron filter, then, add the filtered doxycycline solution to the remaining 95 milliliters of sucrose solution, and supply the solution to the mouse instead of drinking water. Check daily and replace immediately if the solution becomes cloudy as this indicates bacterial overgrowth.

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