- Plate an appropriate number of cells on a gelatinized 60 mm
dish and grow until confluent, feeding daily. Cells should be
passaged without antibiotics prior to submission.
- Feed with cES cell media 1 – 2 hours prior to
- Rinse 2 times with 5 ml PBS.
- Add 2 ml ES cell trypsin.
- Incubate at 37°C for 3 - 5 minutes.
- Counter with 4 ml cES cell media.
- Pipette up and down until colonies are broken into single cell
- Transfer total volume of cells to a 15 ml conical tube.
- Count cells. Cell count is not critical, but it must be
noted if there are more than 5 x 107 cells/ml.
- Centrifuge at 1000 rpm for 5 minutes. Aspirate media.
- Re-suspend pellet in 500 µl PBS.
- Split volume between two 1.5 ml eppendorf tubes.
- Label with “Rodent PCR Panel”, Investigator, Construct, Clone,
Passage Number, Date.
- Store at -80°C.
- Contact DAC Comparative Pathology Lab and arrange for a time to
drop off the cell samples.
Vials will need to be delivered on dry ice after making
arrangements for submission.