The Transgenic Core Facility creates transgenic mice in C57BL/6 and FVB inbred strains, B6CBAF1, B6C3F1 and B6D2F1 hybrid strains, as well as CD1 outbred strain. Contact the facility if you wish to use another strain. We inject standard transgenic DNA constructs as well as larger sized DNA including PAC, BAC and YAC DNA constructs. Services include:
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Perform microinjection and egg transplantation.
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Microinject up to 250 eggs for standard construct (DNA < 20 Kb), and up to 450 eggs for PAC, BAC, and YAC constructs.
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Produce at least three transgenic founders per construct.*
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Wean and transfer the potential transgenic founders to the investigator at 3 weeks of age.
*While more than two transgenic founders are usually produced from each construct, the number varies by DNA construct. If fewer than three founders are produced, the Transgenic Core Facility will inject an additional 100 -150 eggs at no charge. If there are still less than three founders after a second round injection, new DNA needs to be made by the investigator and an additional fee will be charged for re-injection. For large sized DNA, the facility cannot guarantee full-length transgenic founders.
Blastocyst/Morula Microinjection (Download PDF Form)
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Perform mouse blastocyst/morula microinjection and embryo transplantation.
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Microinject 2 - 3 ES cell clones for each construct.
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Produce at least 3 highly chimeric mice.*
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Transfer the chimeric pups to the investigator at 3 weeks of age.
*Chimeric mouse production and germline transmission depend entirely on the pluripotency of the ES cells. If fewer than 3 highly chimeric mice are produced, the Transgenic Core will inject an additional 50 blastocysts/morula at no additional charge. If there are still less than 3 highly chimeric mice, new targeted ES cell clones need to be made by the investigator and an additional fee will be charged for re-injection.
New Mouse ES Cell Line Derivation (Download PDF Form)
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We have successfully derived multiple mutant mouse ES cell (mESC) lines in C57BL/6J, C57BL/6 X C3H/He, and C57BL/6 X DBA/2 backgrounds.
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We will collect blastocysts from the mice provided by the investigator, and isolate, expand mESC colonies.
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Determine sex of new mESC line by PCR.
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Transfer 5 vials of ES cells per line with passage 5 or lower.
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Perform tail biopsy and prepare mouse tail DNA.
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Perform PCR analysis using the primers and method provided by the investigator.
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Transfer the transgenic founders to the investigator.
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Perform enzyme digestion of plasmid DNA and electrophoresis.
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Excise the DNA fragment from agarose gel.
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Purify the DNA fragment for microinjection.
Cryopreservation of Mouse Embryos (Download PDF Form)
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Superovulate female mice and mate with male mice.
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Collect and cryopreserve at least 300 embryos per strain.
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Store in two liquid nitrogen tanks at the facility for three years.
Recovery of Cryopreserved Mouse Embryos (Download PDF Form)
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Superovulate and mate the mice provided by the investigator.
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Collect and transplant fertilized eggs to psudopregnant foster mothers.
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Transfer the pups to the investigator at 3 weeks of age.
