Procedures

For pronuclear injection, investigators provide the DNA construct to the Transgenic Core. The core will:

• Gel purify traditional transgene constructs.
• Harvest single cell embryos.
• Microinject the embryos with the transgene or CRIPSR construct.
• Transplant the microinjected embryos into pseudo-pregnant females.
• Perform cesarean sections as required.
• Provide the resulting pups at weaning age to the investigator for analyses.

The core guarantees the generation of 10 births or one founder (whichever comes first) when using C57BL/6. embryos (historic average is 14 births per round).

The core reserves the right not to take on projects with low chance of success. The core provides C57BL/6 mice. If an investigator needs other strains, he or she will need to pay for them.

For sperm cryopreservation with verification, investigators provide a minimum of two male mice. The core will:

• Harvest sperm from the donor
• Produce 20 straws of frozen sperm
• Verify successful freezing via in vitro fertilization using the thawed sperm and donor eggs from the relevant strain of mice; verification entails development of 2-cell embryos following in vitro fertilization

The verification is the guarantee for this service.

For sperm cryopreservation without verification, investigators provide a minimum of two male mice. The core will:

• Harvests sperm from the donor
• Produce 20 straws of frozen sperm

The core guarantees mobile sperm upon test thawing. Cryopreservation activities are based on a good faith agreement between the investigators and the core. Thus far, the core has not encountered any difficulties in the recovery of cryopreserved samples which it has generated. However, we cannot guarantee that all samples can be recovered following long-term storage.

For in vitro fertilization and embryo transfer, investigators provide either frozen sperm samples or a minimum of two living mice. The core will:

• Thaw the frozen sperm or harvests fresh sperm from the donor
• Performs in vitro fertilization
• Cultures embryos to 2-cell stage
• Transplant the 2-cell embryos into pseudo-pregnant females
• Performs Cesarean sections as required
• Provide the resulting pups at weaning age to the investigator for analyses.

If no births are obtained, a second round (optimized based on the results from the first round) will be performed for no additional charge. Subsequent efforts will require additional billing.

For embryo thaw and transfer, investigators provide the frozen embryos. The core will:

• Transplant the 2-cell embryos into pseudo-pregnant females
• Perform Cesarean sections as required
• Provide the resulting pups at weaning age to the investigator for analyses

If no births are obtained, a second round (optimized based on the results from the first round) will be performed for no additional charge. Subsequent efforts will require additional billing.

Cryopreservation activities are based on a good faith agreement between the investigators and the Transgenic Core. Thus far, the core has not encountered any difficulties in the recovery of cryopreserved samples which it has generated. However, we cannot guarantee that all samples can be recovered following long-term storage.