Recommended control:Human wild-type HeLa cell line (ab255448). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.

Cryopreservation cell medium:Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.

Culture medium:DMEM (High Glucose) + 10% FBS

Initial handling guidelines:Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80 C. Storage at -80 C may result in loss of viability.

1. Thaw the vial in 37 C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmedculture medium, wash vial with an additional 0.8 mLculture medium(total volume 10 mL) to collect remaining cells, and centrifuge at 201x g(rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmedculture mediumand count using a haemocytometer or alternative cell counting method. Based on cell count, seed cells in an appropriate cell culture flask at a density of 2x104cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules.
4. Incubate the culture at 37 C incubator with 5% CO2. Cultures should be monitored daily.

Subculture guidelines:

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