Antibiotic (Hygro) Colony Selection
Mammalian vector V0200 (BD002 Backbone) has selection markers for Ampicillin (bacterial cell selection)and Hygromycin (mammalian cell selection) antibiotics.
Transgenic Cell Maintenance
| Cell Type || Genotype || Medium || Supplements|
| U2OS (adherent) || Flp-in T-REx, hygro-transfected || McCoy's 5A || 10% FBS, 1 % pen/strep || 15 ug/mL blasticidin, 60 μg/mL hygromycin
| U2OS (adherent) || Flp-in T-REx, hygro-transfected || McCoy's 5A || 10% FBS, 1 % pen/strep || 15 ug/mL blasticidin, 200 μg/mL hygromycin
- Treat U2OS transfected cells with 200µg/mL of media for 5 days.
- Cells were happy with new media for the first 2 days and only two small colonies remained on the edge of the plate after 6 days.
- After 6 days growth, cells were again treated with Hygromycin-free media (U2OS media(p/s)).