ES Cell Targeting
Services provided by the facility
- The gene-targeting facility will electroporate DNA provided by the investigator into embryonic stem cells (ES cells) and perform either positive or positive-negative selection.
- ES cells lines provided:
- E14 (129/Ola embryonic stem cell)
- B6-albino (C57BL/6J albino embryonic stem cell)
- CJ7 (129S1/Sv embryonic stem cell)
- W4 (129S6 embryonic stem cell)
- Genomic DNA from surviving clones will be returned to the investigator to assay for targeted clones. Guaranteed 100+ clones for positive-negative selection and 300+ clones for positive selection or perform up to 3 electroporations for each targeting construct.
- The basic service will expand and freeze all "positive clones" and karyotype 4 of these lines for the set price listed below. Any additional expanding and/or karyotyping of clones will be subject to an additional charge listed below.
Responsibilities of the investigator
- Discuss experimental design with Melissa Larson (TGIF director) before beginning a project.
- Provide a map of the construct including size, regions of homology, selection strategy, enzyme sites, and oligo binding sites.
- Purify the DNA targeting vector according to protocols provided by the facility. We’ll need 50ug of purified, linearized DNA at ~1ug/ul.
- Provide gel documentation of the purity and concentration of the DNA target.
- Develop a reliable genotyping assay (long PCR, Southern, or both) before submission of DNA to the facility.
- Submit “Requisition Form” for services to be rendered.
- Storage of frozen ES cells for three months while the DNA is being analyzed by the investigator.
Fees for ES Cell Targeting service
- $4400.00 ES cell targeting (includes expansion and karyotyping of screened, selected ES cells, see above for limit).
- $220.00 additional ES cell karyotyping.
- $330.00 additional ES cell expansion.
- Service Request Form for ES Cell Targeting
Click here for the requisition form for Gene Knockout in ES cells