Initiate a Gene Targeting Project
Do targeted ES cells and/or knockout mice already exist for your gene? Targeted ES cells generated through the KOMP initiative can be found through the UC Davis repository or International Knockout Mouse Consortium. Targeted clones from any of these ES cell resources can be shipped directly to us for injection. Contact us if you need help selecting clones from these sites.
Knockout lines generated through the KOMP2 initiative can be found through the DTCC consortium or International Knockout Mouse Consortium. Cryoarchived lines from any of these resources can be shipped to our lab for cryorecovery.
Initiating a Project
To initiate a project, you will need to provide the following information:
- A completed Gene Targeting Request Form submitted to the lab; Contact us
- Purified electroporation DNA: ~100µg total, linearized, purified DNA prepared at a concentration of at least 2µg/μl sterile TE
- Evidence of a robust screening assay for analysis of homologous recombination in selected clones (it is incumbent up on the investigator to develop a sensitive screening assay and to demonstrate the ability of probes to detect control (wild-type) DNA in 2-5µg genomic DNA isolated from a 96-well plate (protocol); 96-well practice plates are available upon request; contact us to request practice plates
TTML staff provides guidance and advice on targeting vector design, screening strategies and data interpretation.
Once the completed request form and purified DNA is received, we will:
- Review your screening assay
- Verify DNA concentration by fluorometry and assess targeting vector integrity by gel electrophoresis; DNA of inadequate concentration or purity will be returned
- Schedule electroporation