Welcome to BWH Transgenic Core

Gene Targeting Application Form Services & Fees Technical Contacts Address
　
　

Gene Targeting

 　
　

Our ES facility will generate targeted ES cell lines with KO constructs provided by investigators. Upon receipt of the constructs and application form, electroporation will be scheduled. The Core Director, Dr. Arlene Sharpe will prioritize the service requests according to the difficulty of the project and work load.

 

1. Transfection of your linearized DNA construct into ES cells, J1 (129/SV) and Bruce 4 (C57 BL/6J).

2.  After electroporation and selection, clones will be picked, cultured in multi-well plates and replicated. The clones will be frozen to be used to expand positive clones.

3. The investigator will be notified and given DNA isolated from from each clone suitable for screening.

4. Since storage of frozen cells in multi-well dishes is not optimal, it is strongly suggested that researcher return the screening results to the core within 4 months of receipt of the DNA samples.

5. After screening the clones, Up to six ES positive clones will be expanded and frozen down.

6. After expansion, the investigator will then receive DNA from each clone for reconfirmation of targeted recombination. After reconfirmation, the best growing clones will then be scheduled for injection into blastocysts.  Chimeric mice will be given to the investigator.