363+1336 gateaway transfection on 9-14-2012
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| No. |
ID |
Plasmid Name |
miniprep | Lipofectanine 2000 | ||
| 1 |
393+1336-1 |
pDonr221_EGFP+pCMV5-Fc(3C)-Dest-WPRE |
15ul | weak green | ||
| 2 |
393+1336-2 |
pDonr221_EGFP+pCMV5-Fc(3C)-Dest-WPRE |
15ul | weak green | ||
| 3 |
393+1336-3 |
pDonr221_EGFP+pCMV5-Fc(3C)-Dest-WPRE |
15ul | weak green | ||
| 5 |
393+1336-5 | pDonr221_EGFP+pCMV5-Fc(3C)-Dest-WPRE |
10ul |
weak green | ||
| 6 | 393+1336-7 | pDonr221_EGFP+pCMV5-Fc(3C)-Dest-WPRE |
10ul |
weak green |
||
| 7 | 393+1336-8 | pDonr221_EGFP+pCMV5-Fc(3C)-Dest-WPRE |
10ul |
weak green |
||
| 8 | 529 | pLVX-mIL10-IRES-EGFP |
0.5ul |
strong green |
12 well plate transfection (293T cells):
Purpose: want to check the gateaway plasmid expression in the 393 plus 1336 plasmid using 293T cells.
1ug DNA plasmid (miniprep) and 4 ul lipofectamine. Observed weak GFP at very late time 3rd day . For clone 529, transfection efficiency as expected
Conclutsion: weak GFP expression with Lipofectamine 2000. Can't tell which plasmid has better expression.