Gene Targeting Service
Gene targeting service is outsourced to UMich transgenic animal model core:
The lab can begin a new project by obtaining the genomic clone of the investigator’s gene, characterizing it, and constructing the targeting construct.
Accession number or clone
The core can consult with investigators in the design of the targeting construct, and screening strategy.
Following targeting construct synthesis and validation, the core will prepare ES cells and feeder cell dishes, electroporate the cells with the investigator’s targeting construct, apply selection conditions to the cultures, pick, expand and freeze approximately 200 clones, isolate DNA samples from duplicate clone cultures, and analyze for targeting using a PCR and Southern blot strategy.
Once appropriately targeted ES cell clones have been identified, they will be expanded, tested by karyotyping and for mycoplasma, and the GEMM will schedule blastocyst injections with the desired number of clones in consultation with the investigator.