Fees for Service

The UTSW Transgenic Technology Center (TTC) on ND11.223 injects the DNA into fertilized mouse eggs and transfers a minimum of 300 into recipients. The Center cares for the pregnant females and their litters until two weeks after the mice are born. At this time the animals are transferred to the Investigator.

Microinjection                $1,700 -C57Bl/6       $1,700 -B6SJLF1        $1,700 - FVB

Guarantee Policy

In exchange for the fees charged for services, the Center warrants that it will produce at least 3 transgenic founders. We do not guarantee high copy number of the transgene, germline transmission or expression of the transgene. The guarantee is only extended to Investigators whose DNA preparations fulfill the criteria summarized below.

Investigator Requirements

1.  The Investigator generates DNA construct and completes all forms prior to contacting the Center. THE  FOLLOWING REQUIREMENTS MUST BE MET IN ORDER TO BE PLACED ON THE WAITING LIST.

a) Properly prepared DNA construct (See guidelines for preparation of transgenes #3)
b) IDR for the appropriate cost
c) Evidence of a screening strategy (Southern blot, dot blot, PCR)
d) Conventional Transgenic Services Request Form

2. The Investigator contacts the Center staff (x 56287), to reserve a date for microinjection.

3.  The guarantee policy only applies if all of the following criteria are met.  If the client wants to have a DNA sample injected that does not fulfill all of these criteria, the guarantee will not apply.

4. The Investigator prepares DNA from the tails of the putative transgenic mice and analyzes for successful integration of the transgene by PCR, dot blot, or Southern blot within 3 weeks of receiving mice. In the event that there are no transgenic founders, the Center will repeat the microinjection procedures at no charge. This request MUST come within 3 weeks of receiving the animals. The Center will not be responsible for false positives that may result from screening by PCR. A repeat of the microinjection/transfer procedure will require a new DNA preparation and review of the client's screening data by Center staff.

a) We recommend that DNA for microinjection (between 1 and 20kb) be prepared by EluTip-D (VWR Catalog #28151-080). The detailed protocols for this method is available from Center Staff. Note that DNA prepared by QiaQuick columns will not be accepted.

b) A photograph of a gel that demonstrates the quality of the transgene DNA fragment and clean separation from vector DNA must be provided.

c) The DNA must be provided at a minimum concentration of 50 ng/µl in Injection buffer (10mM Tris, pH 7.4; 0.25mM EDTA). A minimum of 60 µl of this DNA solution must be provided to the Center.

d) Samples must have an OD 260/280 ratio of 1.7 or better and a scanning profile indicative of high quality DNA, as measured on the Center’s spectrophotometer. To avoid delays, it is advised that the client quantify the sample prior to submission to the Center. The Center will quantitate the DNA sample by flourometry prior to injection.

e) If the transgene to be injected is derived from a P1, BAC or YAC vector, please contact the Center staff to discuss protocols for DNA preparation.

Additional Services

Isolation of the transgene from the parent plasmid and preparation for microinjection:

The Center will isolate the transgene by restriction digestion, gel electrophoresis and DNA elution. The cost for this service is $400. The requirements to have this done by Center staff include providing 60µg of plasmid DNA, a restriction map of the plasmid detailing the relevant digests needed to liberate the transgene from the vector backbone, and the approximate size of the transgene and vector.

Time Line for Generation of Conventional Transgenic Mice