Cas9 Expressing HT1080 Cell Line | T3260
- 5 to 7 Days Shipment
abm | Cas9 Expressing HT1080 Cell Line | T3260
HT1080 stably expressing Cas9
Thaw entire contents into an appropriate T25 flask as specified in the Propagation instructions.
1. Pre-warm complete media in a 37°C waterbath.
2. Remove the cryopreserved vial from the liquid nitrogen storage tank.
3. Thaw the cells quickly by placing the lower half of the vial into the 37°C water bath and remove after 60 seconds. There should still be a few ice crystals left after thawing. It is important not to over-thaw the cryovials as the presence of DMSO is toxic to the cells.
4. Re-suspend the cells in the vial and transfer the cell suspension into a 15mL sterile conical tube containing 5mL complete media.
5. Centrifuge cells at 200x g for 3 minutes to pellet.
6. Aspirate out the media, leaving cell pellet undisturbed.
7. Re-suspend pellet in fresh culture medium and plate in new culture vessel.
8. Incubate cultures at 37°C, 5% CO2.
The base medium for this cell line is Prigrow III medium available at abm
1) Puromycin selection
2) Western Blot analysis for Cas9 transgene expression
10⁶ cells/1.0 ml