Electroporation
Vetted tissue culture techniques using quality tested ES cell lines and companion reagents are critical in maintaining cell health and viability through the electroporation, selection, and expansion process. Embryonic stem cells (ES) are known to be a very sensitive reagent and are at the core of the production process of gene targeted animals. iTL’s extensively validated cell lines are regularly quality tested and have produced thousands of chimera mice capable of transmitting the ES cell genome to offspring.
Two industry standard cell lines, C57BL/6 and 129svev have provided our customers with a wide range of successes and genetic background purity. Our proprietary Hybrid 129svev/C57BL/6 cell line was developed to combine the strengths of the C57BL/6 and 129svev lines and has resulted in a more robust and efficient ES cell line over the past five years. In May of 2010, we introduced a BALB/c cell line to provide our customers with an expanded series of custom mouse model options.
Once your targeting vector has been complete, it is electroporated into one of these highly efficient ES cell lines. These are then placed under drug selection, and resistant clones are selected and cultured.
Full-service Eletroporation
For customers working with us on full-service mouse model production, the resistant clones are selected and cultured until identification and confirmation of positive clones is achieved.
Component Service Electroporation
Customers utilizing our expertise through component services, in which the customer provides their own targeting vector, iTL delivers to you ES clone DNA samples in a 96- or 32-well plate format. These plate(s) contain extracted DNA for your internal PCR or Southern screening. When the customer identifies a positive clone, the clone identification number is sent back to iTL for continued expansion until your desired clone colony size is achieved.
