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Transgenic and Gene Targeting Core

Embryonic Stem Cell Targeting


The gene-targeting facility will electroporate investigator-provided purified DNA (gene targeting vectors) into embryonic stem cells (ES cells) and perform either positive or positive-negative selection to isolate recombinant ES cell clones. Please contact the Core at Makeamouse for more information or visit the ES Cell Strains information sheet for list of available cell lines.

  1. Investigator purifies the DNA vector
  2. Please provide 30 µg of linearized DNA at a concentration of 1µg/µl. The DNA should be free of chloroform, phenol, ethidium bromide, alcohol and salts. See the recommended DNA preparation protocol. Please submit completed ES Cell Service Request form to the Gene Targeting facility as directed on the form. Please also note that we require proof that your DNA preparation meets our quality requirements (see our protocol DNA Preparation for Gene Targeting; contact Makeamouse for details)
  3. Core purifies the DNA vector; Please contact the gene targeting facility to discuss requirements.

Once targeted ES cell clones have been identified, confirmed by southern analysis or PCR analysis, and passed karyotyping analysis, TGTC will schedule blastocyst injections. Please complete the embryonic stem cell injection service form and submit it to the TGTC through Makeamouse.

 


 

Note:
All communications must be routed through the Makeamouse@ucdenver.edu mailbox. Briefly state your questions or concerns. Your email will be routed to appropriate Core personnel.