1. What do I need to do to have a transgenic or knockout mouse made?

You must have a current approved protocol with the UAB IACUC before the start of your project. 

Please fill in and submit the service agreement form and provide your protocol and account numbers. 

You will need to prepare a concentrated sample of your DNA construct, and provide a sequence file along with a gel image of a sample of your purified fragment (please include a DNA ladder on the gel).  

In the case of ES cell injection, you will need to submit a frozen vial of cells for the Transgenic Core to thaw as the injection schedule permits. We have a system set up for direct shipment of ES cells to our facility from KOMP, EUCOMM, or NORCOMM.

Note: Place your order as soon as possible since there is a considerable backlog before cells will be processed and can be sent out.

Please notify the facility when you expect to deliver the ES cells or the DNA construct so that someone is available to meet you.

2. How long will it take to obtain the transgenic or knockout mouse?

The amount of time will vary according to the nature of the project and the current project load of the facility.

All requests are processed in the order they are received. The minimum time required between the start of a project and the delivery of pups is 5 weeks. The average turnaround time is 7 weeks.

3. How much will it cost?

Current prices for UAB investigators are displayed on the fees page of the website. These prices are subject to change. External investigators can write to us for pricing information. 

4. How should I prepare my DNA construct?

Please verify the construct's composition by sequencing or restriction mapping. Prepare your plasmid prep and digest the plasmid using restriction enzymes to remove the vector backbone. Run the digest on an agarose gel using TAE (Tris-Acetate EDTA) buffer. Purify your linearized construct from the gel using an appropriate spin-column method (Qiagen QIAquick or QIAEX II, GeneClean, Elutip), following the instructions exactly.

Generally, your construct will not be concentrated enough to use a spectrophotomer to calculate concentration, so you must determine the concentration of your purified fragment by running small aliquots in a gel along with a DNA mass ladder, such as Invitrogen's DNA Mass Ladder and Bio-Rad's Molecular Ruler. The final concentration of the construct should be at least 40 ng/µl, and it should be in water or injection buffer (5-mM Tris-Cl, 0.1-mM EDTA, pH 7.4).

5. Is my transgene too big?

No, cloning and DNA handling issues are the only limits to the size of your construct. If you plan on using BAC or YAC DNA, please notify the facility when you submit your request.  BAC DNA must be handled with extra care.

6. Can you make double transgenic mice?

Yes, multiple constructs can be injected simultaneously. Each DNA construct must be provided separately.  In addition, a previously generated transgenic line can be used as a “background strain” for adding another mutation.

7. How can I follow the progress of my project?

Please send an e-mail and we will respond within 1-2 working days.

8. How many transgenic or chimeric mice will I get?

The facility provides a “best effort” guarantee of

• 150 embryos microinjected for DNA and CRISPR projects or 
• 40 blastocysts microinjected for ES cell injection projects.  

DNA injections for Transgenesis: Typically you will get 2-3 transgenic animals if using the C57BL/6 strain or 3-5 transgenic animals using a hybrid line such as B6XSJLF1 X B6. 
ES cell injections: Typically you can expect 2-5 male chimeras to breed.

9. How can I be sure my ES cell line will go germline?

You cannot, however you can increase your chances by having your ES cells karyotyped before they are used by the facility, or by submitting 1 or 2 extra clones for microinjection. Generally only 60-70% of clones contribute to the germline, therefore the use of multiple clones are recommended.

10. Does the core provide services to customers outside of UAB?

For external customers the contract for services clarifies that the core is providing fee-for-service only, and all materials are the property of the customer's institution.

11. What is the policy of the core for importing or exporting cryopreserved samples?

The policy of the core is that the samples belong solely to the investigator and the core is simply providing services to store cryopreserved samples and/or re-animate the samples into live animals.  However, any animal model or derivatives from that model created at UAB are the real property of UAB, and we can not ship the samples (mice and/or cryopreserved sperm or embryos) without having been given authorization that is granted via a Material Transfer Agreement (MTA).