Q: Where is the Tulane Transgenic Core Facility located?
A: The facility is located on Tulane's Uptown campus in Percival Stern Hall, 6400 Freret Street.
Q: What does the Tulane Transgenic Core Facility do?
A: The facility provides the following services for mice: pronuclear injection of DNA, injection of gene targeted ES cells into blastocysts, CRISPR/Cas9 injection, rederivation to convert the health status of mice, in vitro fertilization and cryopreservation. The facility can also provide CRISPR/Cas9 services in rats.
Q: What do I need to do to have a transgenic or knockout mouse made?
A: You must have a current approved protocol with the university IACUC before the start of your project. Please fill in and submit the Request for Service form and provide your protocol and account numbers. You will need to prepare your DNA or ES cells for injection. For CRISPR/Cas9 you will need to purchase crRNA from IDT. The core provides the tracrRNA and the Cas9 protein. You must contact your animal facility's supervisor in order to apprise them of your intent to import mice from the Tulane Uptown Vivarium, as health information or quarantine may be required. You will need to provide a picture of a gel with a sample of your purified fragment and a DNA ladder. Please notify the facility when you expect to deliver the ES cells, RNAs, or the DNA construct so that someone is available to meet you.
Q: How long will it take?
A: The amount of time will vary according to how busy the facility is. All requests are processed in the order they are received, but please have your DNA construct or ES cells ready before you submit your request. The minimum time required between the start of a project and the delivery of pups is 8 weeks.
Q: How much will it cost?
A: Current prices are shown in the table below. The School of Science and Engineering supports the facility, thus its members get a discount.
|Thaw Frozen Embryos
Q: How should I prepare my DNA construct?
A: Please verify the construct's composition by sequencing or restriction mapping. Prepare your plasmid prep and digest the plasmid using restriction enzymes. 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. The final prep of the construct should be at least 50 µl at 50 ng/µl, and it should be in water or injection buffer (5-mM Tris-Cl, 0.1-mM EDTA, pH 7.1).
Q: Is my transgene is too big?
A: 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.
Q: Can you make double transgenic mice?
A: Yes, multiple constructs can be injected simultaneously. Each DNA construct must be provided separately. Please contact Ann Mullin if you want to use multiple guide RNAs in a CRISPR/Cas9 project.
Q: How can I follow the progress of my project?
A: Just send an e-mail to email@example.com and you will receive a response within 1 working day.
Q: How many transgenic or chimeric mice will I get?
A: The facility guarantees at least three pups will be chimeric or transgenic. If there are no transgenic pups produced from 3 injection sessions, you may be asked to provide a new DNA sample.
Q: How can I be sure my ES cell line will go germline?
A: You cannot, however you can increase your chances by having your ES cells karyotyped before they are used by the facility.
Q: How many transgenic and knockout mice has Tulane Transgenic Core Facility generated?
A: As of January, 2020, the facility has produced 302 transgenic mice from 65 different DNA constructs, 163 knockout mice from 23 ES cell lines, created 96 genetically modified mice using 16 CRISPR projects, and 16 genetically modified rats from 1 CRISPR project, and has cryopreserved 64 mouse lines.
Q: What if there are no transgenic or chimeric pups?
A: The facility guarantees at least 3 pups, so the procedures will be repeated. However, investigators may be required to provide a new DNA prep or ES cell clone or pay for additional animals.
Q: Can you provide services for investigators who are not part of the Tulane University?
A: Yes, the facility will work with investigators from other universities in Louisiana who collaborate with Tulane researchers. There are additional charges for investigators outside the Tulane system.
Q: How did the Tulane Transgenic Core Facility get started?
A: The facility received its start-up funding from Tulane University's Wall fund in 2001 and an enhancement grant from the Louisiana Board of Regents. In 2003 rederivation capabilities were added and in 2009 the facility received funds to add cryopreservation. In early 2017 CRISPR/Cas9 began being offered in mice and in 2018 we added CRISPR/Cas9 in rats.