- pEXPR: Hef1a-eBFP2 - in freezer
- pEXPR: Hef1a-L7ae - in freezer
- pEXPR: Hef1a-2x Kturn-eYFP - with Nelson
EXPERIMENTS
- Hef1a eBFP2 & Hef1a 2x Kturn eYFP (positive control)
- Should be yellow and blue
- Hef1a eBFP2 & Hef1a 2x Kturn eYFP & Hef1a-L7ae (negative control)
- Should be blue (minimal yellow)
- pEXPR: Hef1a-eBFP2 - in freezer
- pEXPR: Hef1a-Gal4Vp16 - possibly with Jin? (get in contact with him)
- pEXPR: Hef1a-TAL14 - Brian has, in pENTRY form
- pEXPR: 5xUAS/4xTAL14bs-eYFP - getting from Samira (but missing UAS)
EXPERIMENTS:
- Hef1a-eBFP2 & 5xUAS/4xTAL14bs-eYFP
- negative control (minimal yellow)
- Hef1a-eBFP2 & Hef1a-Gal4VP16 & 5xUAS/4xTAL14bs-eYFP
- positive control (hopefully linear BFP/YFP)
- Hef1a-eBFP2 & Hef1a-Gal4VP16 & Hef1a-TAL14 & 5xUAS/4xTAL14bs-eYFP
- Should be blue (minimal yellow)
n-WELL PLATE PROPORTIONS:
| 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 |