Transgenic and Knock-Out MousePolicies & Procedures
Transgenic and Knock-Out Mouse Core service charges are as follows. See below for transgenic and knock-out mouse production procedures.
Transgenic and Knock-Out Mouse Core Service Charges |
|
|---|---|
| Service | Pricing |
| Pronuclear injection | $5,400 |
| ES Transfection | $5,605 |
| Blastocyst Injection | $5,408 |
| F0 Pronuclear injection | $2,700 |
| Single cell line injection | $2,704 |
| Tetraploid aggregation | $1,000 |
| IVF | $1,200 |
| Cryo pilot study | $1,500 |
| EX tfx with HAT selection | $850 |
| Embryo cryo | $500 |
| Sperm cryo | $375 |
| Thaw and transfer | $500 |
| Cryo maintance fee | $50/line/yr |
Procedures
- For transgenic mouse production, investigators provide the DNA construct to the core. The core then:
- harvests single cell embryos,
- microinjects the embryos with the DNA construct,
- transplants the microinjected embryos into pseudo-pregnant females,
- performs Cesarean sections as required, and
- provides the resulting pups at weaning age to the investigator for analyses.
- For knock-out mouse production, the core provides three types of services:
- For ES cell transfection, the investigator provides the core with a targeting vector. The core then:
- transfects ES cells with the targeting vector,
- selects the resulting transfected clones,
- generates frozen stocks of the clones, and
- provides the investigator with DNA samples from each of the clones for molecular analyses.
- For blastocysts injections, the investigator provides (or identifies) the desired ES clone. The core then:
- recovers and amplifies the ES cell line,
- provides the investigator with cells for additional molecular and cell biologic analyses,
- generates back-up frozen stocks,
- injects the cells into blastocysts,
- transplants the injected blastocysts into pseudo-pregnant females,
- performs Cesarean sections as required,
- culls the resulting chimeric animals, and
- either provides the chimeric pups at weaning age to the investigator for analyses or uses the pups for rapid germ-line breeding.
- For rapid germ line breeding, core personnel:
- rotates female mice with the chimeric males,
- replaces plugged females until a total of 10 pregnancies are culled,
- screens the resulting pups for germ line transmission of the ES cell genome (determined via coat color),
- provides tail biopsies to the investigator for molecular confirmation of germ line transmission, and
- provides the resulting heterozygous animals at weaning age to the investigator for analysis.
- For ES cell transfection, the investigator provides the core with a targeting vector. The core then: