Services and Pricing

Pronuclear DNA Microinjection

This service provides an efficient means of introduction foreign DNA, usually plasmid or BAC derived fusion genes, into the mouse genome. DNA is microinjected into the pronucleus of 0.5 day fertilized mouse embryos. Surviving embryos are transferred to the oviduct of a pseudopregnant recipient and the animals are housed under the protocol number of the investigator but husbandry is provided by the TMESCSR and DAC. The gestation period of the mouse is 19 to 21 days. Pups are born, and will be weaned, tailed and numbered by core personnel at 3-4 weeks of age. The investigator will need to perform DNA isolation and analyze the DNA by PCR or Southern blotting to identify transgenic founders.

Several factors influence the production of transgenic mice. Of utmost importance for successful outcome is the quality and size of the DNA injected, thus no guarantee is made by the Transgenic Mouse/Embryonic Stem Cell Shared Resource for the number of transgenic mice produced. However, provided that the DNA has been prepared in the recommended manner, our records indicate that 10 to 20% of the pups born will usually carry the transgene.

Experimental Flow Chart:

1. PI submits frozen DNA and appropriate service form to TMESCSR for microinjection
2. Service forms are reviewed and approved by the Co-Directors of the facility, DNA is quantitated, quality controlled, and diluted by Tg Mouse/ESC staff
3. Injections are scheduled, mice are ordered
4. Injections are performed, surviving embryos are transferred to pseudopregnant females
5. Pups born 19 to 20 days post injection
6. Pups weaned, tailed and ear punched by TMESCSR staff at 3 weeks of age
7. PI identifies transgenic founders from tail DNA using PCR or Southern blotting
8. PI notifies TMESCSR staff about transgenic founders identifying individual mice
9. Non-transgenic littermates are serology tested at 6 weeks of age
10. DAC staff review serology results, approve movement of mice, and transfer transgenic founders to PIs mouse housing room at 7 weeks of age