TRANSFECTION PLANS
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| Name | Location | Concentration | | MAV1212-hEF1a-eBFP2 | TC Room | Midi prepped | | pEXPR:hEF1a-L7ae | TC Room | Midi prepped | | pEXPR: CAG-2xKturn-eGFP (wt) | TC Room | Midi prepped | | pEXPR: CAG-exKturn-eGFP (mut) | TC Room | Midi prepped |
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)
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| Name | Location | Concentration | | pEXPR: hEF1a-eBFP2 | TC Room | Midi prepped | | pEXPR: hEF1a-Gal4VP16 | Our -20 | Verified, colony being picked for midi prep | | pEXPR: hEF1a-TAL14 | Our -20 | Being midi prepped | | pEXPR: TAL14-mKate | TC Room | Midi prepped |
EXPERIMENTS: - hEF1a-eBFP2 & 5xUAS/4xTAL14bs-eYFP
- negative control (minimal yellow)
- hEF1a-eBFP2 & 5xUAS/4xTAL14bs-eYFP & hEF1a-Gal4VP16
- positive control (hopefully linear BFP/YFP)
- hEF1a-eBFP2 & 5xUAS/4xTAL14bs-eYFP& hEF1a-Gal4VP16 & hEF1a-TAL14
- Should be blue (minimal yellow)
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| name | Repressor co-transfection experimental designs |
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NEED TO TEST
Goal: Test the strength of different repressors
| Name | On | Off | Inducible |
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| L7ae | Gateway Hef1a-2x Kturn-eYFP with pENTR_L1_L7ae_L2 | Just Hef1a-2x Kturn-eYFP | rtTA & DOX? |
| TAL14 | Gateway Hef1a-2x Kturn-eYFP with pENTR_L1_TAL14_L2 | Just Hef1a-2x Kturn-eYFP | rtTA & DOX? |
Notes:
2x-Kturn-eYFP from Nelson