The University of Queensland
Brisbane, Queensland 4072
Australia

Email:e.williams@imb.uq.edu.au
Ph +61 7 33462276
Fx +61 7 33462101

©TASQ 2005
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Microinjection into the pronucleus (Pronuclear Injection, PNI) has been the most used technique for transgenic animal production. Microinjection involves the mechanical introduction of a purified double stranded DNA sequence into the pronucleus of a fertilized mammalian oocyte leading to the integration of the sequence (transgene) within the existing genetic sequence. This integration means that the animal is born with a copy of the new sequence in every cell, making it an excellent method for studying mammalian growth and pathology.

Purification and DNA Preparation Protocol (MS Word)

Prices

  • $AU 950 / session (internal)
  • $AU 1300 / session (external)

These prices cover:

  • Superovulated and mated BCBF1 donor females
  • Embryo collection
  • Microinjection of DNA construct
  • Pseudo-pregnant females
  • Embryo transfer
  • Consumables
  • Agistment until pups are weaned.

Additional Notes and Recommendations

  • TASQ recommends 2 intitial sessions per construct.
  • The Client must provide linarized DNA in an approved injection buffer with known concentration and a gel picture of DNA and plasmid size. TASQ will perform a final preparation of DNA prior to injection.
  • A TASQ application form for Pronuclear Injection must be completed prior to the commencement of service.
  • Additional agistment costs apply once pups are weaned and for the transport of mice if applicable.
  • AEC and GTR numbers must be supplied prior to commencement of service.
  • Penalty fees apply to late cancellations of Injection sessions.
  • The service being provided is undertaken with no guarantee of the genetically manipulated mouse strain.
  • The generation of this genetically modified mouse strain will not infringe any intellectual property rights.

Pronuclear Injection References

Brinster, R.L. et al (1985) Factors affecting the efficiency of introducing foreign DNA into mice by microinjecting eggs. Proc. Natl. Acad. Sci. 82, 4438-4442.

Gordon, J.W. et al (1980) Genetic transformation of mouse embryos by microinjection of purified DNA. Proc. Natl. Acad. Sci. 77, 7380-7384.

Jaenisch, R. (1988) Transgenic Animals. Science. 240, 1468-1474.