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Protocol for prepping ES cells for the rodent PCR panel by Charles Rivers Laboratories for PCR viral detection and mycoplasma testing.

Record History
Added on February 3, 2012 at 12:56 PM by Skelton, Jennifer
Modified on February 3, 2012 at 1:00 PM by Skelton, Jennifer
Shared with (contributions)
TMESCSR: Protocols Master
  1. 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.
  2. Feed with cES cell media 1 – 2 hours prior to trypsinization.
  3. Rinse 2 times with 5 ml PBS.
  4. Add 2 ml ES cell trypsin.
  5. Incubate at 37°C for 3 - 5 minutes.
  6. Counter with 4 ml cES cell media.
  7. Pipette up and down until colonies are broken into single cell suspension.
  8. Transfer total volume of cells to a 15 ml conical tube.
  9. Count cells.  Cell count is not critical, but it must be noted if there are more than 5 x 107 cells/ml.
  10. Centrifuge at 1000 rpm for 5 minutes. Aspirate media.
  11. Re-suspend pellet in 500 µl PBS.
  12. Split volume between two 1.5 ml eppendorf tubes.
  13. Label with “Rodent PCR Panel”, Investigator, Construct, Clone, Passage Number, Date.
  14. Store at -80°C.
  15. Contact DAC Comparative Pathology Lab and arrange for a time to drop off the cell samples.

Anne Pate


MCN T322

Vials will need to be delivered on dry ice after making arrangements for submission.