ES Cell Targeting

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:
- B6-white (C57BL/6 albino embryonic stem cell)
- CJ7 (129S1/Sv embryonic stem cell)
- W4 (129S6 embryonic stem cell)
- Bruce4
- RW4
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 subjected to an additional charge listed below.

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.

$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.
A yearly storage fee of $50.00 will be charged for 1-50 vials of targeted ES cells, $100.00 for 51-100 vials, and $150.00 for 101-150 vials.
Service Request Form for ES Cell Targeting