Human iPS Cell Gene Targeting

Responsibilities of the Investigator

  • Discuss experimental design with facility directors before beginning your project.
  • Provide a map of the construct including size, regions of homology, selection strategy, enzyme sites and oligo binding sites.
  • Identify a unique restriction site to linearize your plasmid.  Provide gel documentation of your test digest.
  • Provide 100ug of purified plasmid at 1ug/ul.
  • Develop a reliable genotyping assay (long PCR, Southern, or both) before submission of DNA to the facility.
  • Screen picked clones for targeted candidates within one month (during which time the clones are stored at -80o).
  • Confirm genotype of candidate clones by Southern blot.
  • Submit a Request Form in iLabs to initiate the project.

Services Provided by the Facility

  • The gene-targeting facility will linearize your plasmid, electroporate into ES/iPS cells, perform selection and pick clones.
  • ES cell lines provided:
  • iPS cell lines provided: 
  • We guarantee two rounds of electroporations (if necessary) for each targeting construct.
  • Picked clones will be stored at -80o for up to one month, during which time the investigator must screen for candidate clones.
  • The facility will expand candidate clones for freezing and confirmation of genotyping by Southern blot (investigator provides Southern blot confirmation).
  • After confirmation of homologous recombination, the facility will karyotype up to four of these clones.  Any additional expansion and/or karyotyping of clones will be subject to an additional charge.

Fee for Human iPS Cell Gene Targeting Service

TBD

Last modified: Dec 14, 2016
Contact Us

Phone: 913-588-7113

Email: TGIF@kumc.edu

Submit request in iLabs

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