User:Behzad Damadzadeh/Notebook/PcTF Subcloning in E-coli/2013/06/21: Difference between revisions
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''' | ==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''' | |||
{| {{table}} cellspacing=''7'' | |||
|- valign="top" | |||
| '''Cell Type''' || '''Genotype''' || '''Medium''' || '''Supplements<br>(required)''' || '''Maintenance<br>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 | |||
|- | |||
|} | |||
<br> | |||
*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)). |
Revision as of 16:35, 23 July 2013
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06/21/2013Antibiotic (Hygro) Colony SelectionMammalian vector V0200 (BD002 Backbone) has selection markers for Ampicillin (bacterial cell selection)and Hygromycin (mammalian cell selection) antibiotics.
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