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STEP | COMPONENT | 6-well | 24-well | 96-well |
---|---|---|---|---|
1. | Adherent Cells | 0.25-1 x 10^6 | 0.5-2 x 10^5 | 1-4 x 10^4 |
3. | Opti-MEM Medium | 150uL x 4 | 50uL | 25uL x 4 |
Lipofectamine LTX Reagent | 6, 9, 12, 15 uL | 1.5uL | 1, 1.5, 2, 2.5 uL | |
4. | Opti-MEM Medium | 700uL | 250uL | 125uL |
DNA (0.5-5 ug/uL) | 14ug | 5ug | 2.5ug | |
PLUS Reagent | 14uL | 5uL | 2.5uL | |
5. | Diluted DNA (with PLUS) | 150uL | 50uL | 25uL |
Diluted Lipofectamine LTX | 150uL | 50uL | 25uL | |
7. | DNA-lipid complex (per well) | 250uL | 50uL | 10uL |
FINAL DNA (PER WELL)** | 2500ng | 500ng | 100ng |
Plasmid (Concentration) | 6-well | 24-well | 96-well |
---|---|---|---|
pEXPR hEF1A:eBFP2 (96 ng/ul) | 13 ul (2 plasmids) 8.7 ul (3 plasmids) 6.5 ul (4 plasmids) | 2.6 ul (2 plasmids) 1.8 ul (3 plasmids) 1.3 ul (4 plasmids) | .5 ul (2 plasmids) .4 ul (3 plasmids) .3 ul (4 plasmids) |
pEXPR hEF1A:l7ae (186 ng/ul) | 4.5 ul (3 plasmids) | .9 ul (3 plasmids) | - |