The facility provides a comprehensive service for the generation of genetically modified mouse models including:
- Design of recombinant DNA molecules for genetic modification.
- Design of screening protocols for mice and ES cell clones (PCR and Southern-blot).
- Generation of gene-targeting vectors by recombineering.
- Help with cloning protocols and reagents required to generate gene targeting and transgenic vectors.
- All cell-culture required for gene targeting in mouse embryonic stem (ES) cells, including expansion, transfection, drug selection, clone picking, clone screening by PCR, and clone expansion/ confirmation (PCR based).
- Microinjection of DNA or ES cells into pre-implantation stage embryos.
- Transfer of manipulated embryos into pseudo-pregnant foster mice.
- Subsequent processing of pups born to pseudopregnant fosters.
The facility has a fully equipped laboratory for cell and molecular biology. Mouse embryonic stem (ES) cell culture is carried out within a dedicated cell culture laboratory. There is a small microinjection suite with inverted microscopes equipped with state of the art micromanipulators and microinjectors, based within the PCB Animal Research Center (PRAAL). The facility has everything necessary for mouse surgery and embryo manipulation, and it is equipped to carry out cryopreservation of mouse germ-line tissue.