Encyclopedia of Experiments
Biological Techniques
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Encyclopedia of Experiments Biological Techniques
CreER-LoxP System-Based Target Gene Inactivation: A Tamoxifen-Inducible Cre Recombinase System for Target Gene Knockout in a Mouse Model

CreER-LoxP System-Based Target Gene Inactivation: A Tamoxifen-Inducible Cre Recombinase System for Target Gene Knockout in a Mouse Model

Transcript

Initiate induction of Cre Recombinase 10 to 15 days after tail vein injection, to allow for clearance of non-integrated vectors. Dissolve 10 milligrams of tamoxifen in 40 microliters of ethanol, and incubate at 55 degrees Celsius for 10 minutes with shaking.

Vortex several times until the tamoxifen has dissolved, then add 960 microliters of corn oil, and incubate for 5 minutes at 55 degrees Celsius. After cooling, draw the solution into a 1-milliliter insulin syringe with a 20-gauge needle, then change to a 27-gauge needle.

To inject, first, scruff the mouse by grabbing the neck of the mouse carefully with the thumb and the second finger, fixing the tail between the base of the hand and the fourth and fifth finger, then, intraperitoneally inject 0.1 milliliters of the solution into the left lower quadrant of the abdomen.

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