06/21/2013
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
(required) |
Maintenance
antibiotic(s)
|
| 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)).
|
PcTF Subcloning in E. coli
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