Pronuclear Injection of DNA
Responsibilities of the Investigator
- Obtain IACUC approval for your project before requesting service.
- Discuss your transgene design with the facility directors before initiation of the project. Important: when designing your construct, ensure the transgene sequence is flanked by unique restriction enzyme sites to allow for easy removal of the vector backbone. Isolation of your insert is essential, as inclusion of prokaryotic sequence may interfere with expression of your transgene.
- Provide 25ug of circular plasmid DNA containing your construct at a concentration of 1mg/ml in TE buffer.
- Provide a bacterial plate if submitting a BAC for injection.
- Provide a restriction map and sequence of the construct including size, vector, promoter, gene of interest and/or reporter gene.
- Submit a Request Form in iLabs to initiate your project.
Services Provided by the Facility
- The Transgenic Facility will linearize, purify and quantitate your construct for injection.
- We will use F1 hybrid FVB x C57Bl/6 mouse embryos, unless requested by the user and approved by the facility.
- Genotyping will be provided by the facility to identify transgenic founders.
- The facility will guarantee two founders. Exception: The facility does not prepare YAC DNA, and thus will guarantee only the injection and transfer of 200 embryos.
- We cannot guarantee expression and/or transmission of your transgene.
Feb 12, 2021