Gene Targeting
Providing a complete service to generate genetically altered mouse strains for Babraham Institute research groups and external companiesTechnical services and assistance available to researchers
Design and generation of gene targeting constructs
In silico design and cloning
ES cells targeting
- Simple loss-of-function
- Conditional knockout
- Cre reporter allele
- Other alleles: gain-of-function, point mutation
In silico design and cloning
- Southern screening strategy for ES cells
- PCR screening strategy for ES cells
- Generate maps and sequence files for construct in every step
- Search from database for genomic clones: 129/Sv versus C57BL/6 DNA
ES cells targeting
- Preparation of DNA for electroporation of ES cells
- Electroporation of targeting vector DNA into ES cells
- Selection of targeted ES cells: positive and negative selection
Molecular analysis of ES cells clones
Introduction of ES cells into host embryos to create germline chimeras
DNA pronuclear injection for transgenic mouse generation
- Preparation of genomic DNA from ES cells for Southern blot or PCR
- Screening of ES cell clones
Introduction of ES cells into host embryos to create germline chimeras
- Microinjection of ES cells into blastocysts
DNA pronuclear injection for transgenic mouse generation
- Microinjection of DNA into fertilised oocytes
Enquiries can be made via the facility's 'Contact Us' web page.