1353-1305-1369 transfection on 0914-2012
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| No. |
ID |
Plasmid Name |
conc.(ug/ul) | Lipofectanine 2000 |
| 1 |
1353 |
pCMV5-Fc(3C)-RFP-WPRE |
2.0 | weak red at 50h |
| 2 |
1305 |
pCMV5-Fc(3C)-TagRFP |
1.0 | weak red at 50h |
| 3 |
1369 |
pCMV5-Fc-(3c)-TagRFP-WPRE-oriP |
2.8 | weak red at 50h |
| 5 |
529 |
pLVX-mIL10-IRES-EGFP |
2.8 | strong |
12 well plate transfection (293T cells):
Purpose: want to check the stongest expression in the 1353, 1305, 1369 plasmid using 293T cells.
To avoid totoxity of lipofectamine, use 0.5ug DNA plasmid (made 1:20 dilution) and 2 ul lipofectamine. Change medium after 5 hs transfection. Observed weak RFP at 50 hs on monday. For clone 529, transfection efficiency as expected and 0.5 ug plasmid is less efficient than 1 ug plasmid.
9/21/2012
1369: pCMV5-Fc-(3c)-TagRFP-WPRE-oriP
1353: pCMV5-Fc(3C)-RFP-WPRE
1305 pCMV5-Fc(3C)-TagRFP
--Paul.xu 17:25, 21 September 2012 (EST)
Photo taken on 9/23/2012
Conclutsion: weak RFP expression with Lipofectamine 2000. Can't tell which plasmid has better expression.