*There are a few simple provisos:

1. We will purify the DNA for you. Simply perform a restriction enzyme digest on your cloning vector to liberate 50 ug of the transgene insert from the cloning vector.  NEW: We need you to submit 50 ug of DNA,

2. Run out a few hundred nanograms of DNA on a minigel to determine that the digest went to completion and that the bands are the correct size. Bring a photo of the minigel to submit with your sample. Mark the bands on the photo to show which band should be purified for microinjection.

3. Bring the remainder of the digest (in a final volume of 100 to 150 microliters) to the Core lab and we will purify the DNA for microinjection from the digest. We use the Mo Bio UltraClean™ GelSpin™ Kit for purification of microinjection DNA. Please note, if you want use large DNA fragments such as bacterial artificial chromosomes, that there is a different protocol for the preparation of the BAC DNA for microinjection.

4. Bring a gel photo that shows you have a PCR assay for an endogenous mouse gene such as beta-globin. All DNA samples from potentially transgenic mice should give a positive result with an endogenous gene PCR. This will make sure that no transgenic founders are discarded because the DNA concentration interfered with successful PCR.

5. Bring a gel photo that shows your genotyping PCR  detects the transgene at the single copy level when it is mixed with mouse tail DNA. Bring  your calculations of copy number along with the gel photo. If you need tail DNA to set up the assay, we will give you some.

6. Bring a submission form with your contact information and billing information.

7. Drop off all your materials at the microinjection lab: Room 2526, Building MSRB I.

Transgene constructs are purified,  quantitated, and microinjected into (C57BL/6 X SJL)F2 mouse eggs and surgically transferred to recipients. Other mouse eggs can be used for transgenic production when prior arrangements are made with the Core. Contact Thom Saunders regarding custom mouse strains.

Transgenic mouse production is a fee for service provided by the Transgenic Core. The Core prioritizes all requests for service on a "first-come, first-serve" basis. Your DNA will be added to the microinjection queue in the order that it is received.

comments to: tsaunder@umich.edu