To use the Gene Targeting and Transgenic Facility for DNA injection:

1. Submit a TRANSGENIC MOUSE REQUEST FORM to the Gene Targeting and Transgenic Facility. Include a brief outline of the intended experiment that describes the DNA construct to be injected and the experimental question or objective of the experiment. Investigators must make certain, prior to initiating the transgenic work, that they have DNA screening procedures that will detect a single copy transgene present in the mouse genomic background, and must demonstrate this prior to acceptance of the construct.

2. The Gene Targeting and Transgenic Facility will review and approve the submitted request.

3. We have Institutional Animal Care and Use Committee (IACUC) approval for the necessary manipulations to be performed by our personnel in our facility to generate the transgenic animals.

4. You will need to obtain IACUC approval and a protocol number for your intended experiments using the transgenic mice.

5. Provide the GTTF with a high quality DNA preparation of your construct, restriction digested to separate from all vector sequences. You should provide one appropriately labeled 1.5 ml tube containing 10 micrograms of DNA in a restriction digestion reaction with the enzymes, after an incubation period, and a gel photograph of a similar digest in which you have marked the transgene band with a circle or arrow so that it is clear which band is the transgene.THIS IS A CHANGE FROM PREVIOUS PROTOCOL. We will now run the gel and isolate your transgene DNA for you, using our protocol. This new policy is being instituted to increase the transgene integration rate. The DNA tubes should be accompanied by a copy of your service request form.

6. We will initiate an IDT to transfer funds for the mouse expenses (purchase and perdiems for your injections) when we begin the injection series. This portion of the fee is about half.

7. We will notify you when you should expect to pick up the tail biopsies, and then the mice. You will want to perform a timely and well-controlled analysis of the tail biopsy DNA, so that transgenic animals can be identified, and your cage costs can be conserved.  We now guarantee 3 founders per construct, and therefore require that you submit a copy of your screening results to us, with appropriate single copy detection controls by PCR or Southern blotting.

8. Make arrangements for the care and breeding of your transgenic founders in Vivarium space outside of our facility. Once outside of our facility, mice may not reenter our barrier colony. The form page can be filled out and given to the Vivarium, so they will know to expect your mice.