Electroporation of Cre Recombinase

  1. Prior to initiation of any procedure involving Cre recombinase, a material transfer agreement must be signed and returned to the Core Director.
  2. A positive ES clone will be electroporated with the Cre recombinase plasmid.
  3. Customers will be notified of the electroporation date and the expected date for their receipt of DNA samples (see Timeline for approximate schedule).
  4. After electroporation, approximately two hundred clones will be isolated, cultured in multi-well plates, and tri-plated.
  5. One replicate plate will be placed in frozen storage, one plate will undergo selection, and one plate will be incubated without selection.
  6. DNA will be isolated from selected clones and sent to the customer, at their expense, for confirmation of the recombination event.
  7. Since storage of frozen cells in multi-well dishes is not optimal, it is strongly suggested that customers return the genotype results to our laboratory within four weeks of receipt of the DNA samples.
  8. Upon confirmation of the positive clones, the gene targeting core will thaw five positive clones.