The Unit is fully equipped for

1) Molecular biology procedures such as cloning, mutagenesis etc.
2) Cell culture
3) Microinjection with 2 complete setups for pronuclear and blastocyst injections. The animal house is equipped with individually ventilated cages and is operated under specific pathogen free (SPF) conditions. Mice can be housed under sterile conditions until they can be transferred to the care of the customer.

The following services are provided:

Generation of transgenic mice

DNA constructs are purified and injected into both pronuclei of the FVB/n mouse oocytes. 150-200 oocytes are routinely injected per construct. DNA injected into the pronucleus integrates into chromosome(s) and is transmitted to the germ line. Injections are repeated at no additional cost in case they do not result in positive transgenic animals. Services also include Southern blot analysis of tail biopsy DNA as well as pronuclear injections of BAC DNA constructs.

Generation of conventional/conditional “knock-out”, “knock-in” and “chromosome engineered” mice:

We offer the following services for each step of targeted mice production:
  • Targeting vector design includes bioinformatics analysis and cloning strategy design. For genomic sequences that are difficult to clone we provide alternative strategies.
  • Embryonic stem (ES) cell line generation from the desired mouse lines, including karyotype analysis and characterization of ES cell for the germ-line competence.
  • ES cell electroporation and processing (expansion, analysis and freezing) of individual ES colonies. Screening of ES cells for positive targeting events is performed with the help of nested PCR and Southern-blot analysis. All positively targeted colonies are subjected to karyotype analysis.
  • Blastocyst injections of positively targeted ES colonies into B6D2F1 hybrids, as well as tetraploid blastocysts, with production either chimaeras or 100% ES derived animals.

Re-derivation of mouse colonies

In order to produce infection-free animal models, mouse lines are held in a specific pathogen free (SPF) environment. Embryos can be stored for a long period of time by transferring them from a “contaminated” environment to SPF conditions.

Long-term storage of mouse sperm

Mouse sperm is obtained and frozen with consecutive in vitro fertilization procedures.

Getting started:

The investigator is requested to meet with the facility to staff to discuss specific needs and to design the project.