MMRRC Testing of KOMP ES Cell clones includes development of genotyping protocols, genetic verification at the 3' and 5' ends, vector copy number = 1, presence or loss of Y chromosome, and mycoplasma and MPV testing. Type of genetic testing varies for CSD and Velocigene targets. Approximately 30% of KOMP clones tested fail genetic analysis and will require a new clone be selected for expansion and testing. Based on the average germline rate for KOMP targets, we recommend requesting and injecting 2-3 clones per targeted allele. All mutant ES cell lines are heterozygous for the mutation, meaning they carry one modified allele and one wild type allele. Please note, there is high incidence of mixed cell populations in the CSD targeted cell lines, and while our testing can detect this in most cases, it is not 100%, and there may be a very small percent of non or incorrectly targeted ES cells in the population; therefore we strongly recommend targeting verification of the derived mice with 5’ or 3’ LRPCR and/or Loss of Allele analysis. MMRRC also recommends performing chromosome counts or karyotyping prior to injection of clones.

Estimated timelines: Processing ES cells (including genetic verification) prior to injection or shipping requires 12-16 weeks. Chromosome counting (to determine % euploid cells) requires an additional 4-8 weeks. Microinjection requires an additional 11-15 weeks. Germline transmission testing (breeding of chimera) requires an additional 8-16 weeks. Germline transmission is not guaranteed.

Distribution of this strain requires submission of the KOMP MTA. The MTA should be signed by an authorized official from your institution, generally this is someone in your Technology Transfer or Intellectual Property offices. A link to this form will be provided via email when an order is submitted; the PDF form should be downloaded or the email forwarded to the appropriate institutional representative for completion following directions in the document.