- BACs: Verification of BAC clones by PCR
- Targeting vector: Complete sequencing of targeting vector
- Targeted ES cell lines: Karyotyping, Mycoplasma tests by PCR
- Parental ES cell lines: Regular QC regarding germline capacity
- Genotyping:
- Targeted ES cell lines: Southern blot hybridization using 5' external, 3' external and internal probes
- Mice: PCR, quantitative PCR, Southern blot hybridization
- Blastocyst re-implantation: Transfer of injected blastocysts under sterile conditions
- Recipient Mice: Internal breeding of recipient mice under controlled SPF conditions
- Documentation of different mouse production steps by project management software and Laboratory Information Management System (LIMS).
Mouse Housing and Health Screening
Animal housing takes place in a state-of-the-art SPF-facility. This facility is contained within a multi-barrier unit and serviced by HEPA-filter air treatment. In addition, all mice are housed in Individually Ventilated Cages (IVC). The SPF (Specified Pathogen Free) status is monitored by a health screening program with sentinel animals every 3 months for intercurrent infections (viruses, bacteria, mycoplasma, and parasites) exceeds the Federal European Laboratory Animal Science Association standards (FELASA) for the health monitoring of rodent and rabbit colonies in breeding and experimental units (Laboratory Animals (2001) 36: 20-42). Health monitoring is performed by an independent laboratory (Taconic). All work is performed in accordance with all legal requirements including the German Gene Technology Act and the German Animal Welfare Act.
TECHNOLOGY
