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Experiment #1

GOAL: Test the strength of L7ae

3 incomplete 7/14/14: Seed cells 22 incomplete 7/15/14: Transfect MAV1212-hEF1a-eBFP2 (___ ng/uL), Mutant hEF1a-2xKturn-eGFP (___ ng/uL) and Wild Type hEF1a-2xKturn-eGFP (___ ng/uL) into HEK cells 0 incomplete 7/16/14: Flow Cytometer

 

  • DNA Dilution: ___uL MAV1212-hEF1a-eBFP2, ___ uL Mutant Hef1a-2xKturn-eGFP, ___ uL Wild Type Hef1a-2xKturn-eGFP


24-WELL PLATE TRANSFECTION MAP:

(B)1

  • MAV1212-hEF1a-eBFP2

(B)2

  • MAV1212-hEF1a-eBFP2
////////////////////////

(GM)1

  • Mutant Hef1a-2xKturn-eGFP

(GM)2

  • Mutant Hef1a-2xKturn-eGFP

(GM + B)1

  • Mutant Hef1a-2xKturn-eGFP
  • MAV1212-hEF1a-eBFP2

(GM + B)2

  • Mutant Hef1a-2xKturn-eGFP
  • MAV1212-hEF1a-eBFP2

(GM + B + L)1

  • Mutant Hef1a-2xKturn-eGFP
  • MAV1212-hEF1a-eBFP2
  • hEF1a-L7ae

(GM + B + L)2

  • Mutant Hef1a-2xKturn-eGFP
  • MAV1212-hEF1a-eBFP2
  • hEFa-L7ae

(GWT)1

  • Wild Type Hef1a-2xKturn-eGFP

(GWT)2

  • Wild Type Hef1a-2xKturn-eGFP


(GM WT + B)1

  • Wild Type Hef1a-2xKturn-eGFP
  • MAV1212-hEF1a-eBFP2

(GM WT + B)2

  • Wild Type Hef1a-2xKturn-eGFP
  • MAV1212-hEF1a-eBFP2

(GM WT + B + L)1

  • Wild Type Hef1a-2xKturn-eGFP
  • MAV1212-hEF1a-eBFP2
  • hEFa1-L7ae

(GM WT + B + L)2

  • Wild Type Hef1a-2xKturn-eGFP
  • MAV1212-hEF1a-eBFP2
  • hEF1a-L7ae

Neg Ctrl

  • Dummy DNA
  • With Lipofectamine


Neg Ctrl

  • Dummy DNA
  • With Lipofectamine


Neg/Neg Ctrl

  • No DNA
  • No Lipofectamin

Neg/Neg Ctrl

  • No DNA
  • No Lipofectamine
////////////


RESULT: Brief description of experimental outcome (mean transfection efficiency and implications). Link to results page with data analysis.

Experiment #2

GOAL: Test the strength of TAL14

2 incomplete

Day 1: Seed cells

1 incomplete

Day 2: Transfect Promoter:Gene1 (___ ng/uL - transfection marker), Promoter:Gene2 (___ ng/uL) and Promoter:Gene3 (___ ng/uL) into HEK cells

7 incomplete

Day 4: Flow Cytometer

DNA Dilution: ___ uL

Mutant Hef1a-2xKturn-eGFP

hEF1a-eBFP2, ___ uL

Wild Type Hef1a-2xKturn-eGFP

UAS-GAL4-TAL14-eYFP, ___ uL hEF1a-

L7ae
  • (Eg)x: add _:_:_
    1. __uL Promoter:Gene1-lipid complex
    2. __uL Promoter:Gene2-lipid complex
    3. __uL Promoter:Gene3-lipid complex

    • GAL4VP16, ___ uL hEF1a-TAL14


    24-WELL PLATE TRANSFECTION MAP:

    (B)1

    • MAV1212-hEF1a-eBFP2

    (B)2

    • MAV1212-hEF1a-eBFP2
    		//////////////////

    Neg Ctrl

    • Dummy DNA
    • With Lipofectamine
    • MAV1212-hEF1a-eBFP2
    • 5xUAS/4xTAL14bs-eYFP
    • MAV1212-hEF1a-eBFP2
    • 5xUAS/4xTAL14bs-eYFP
    		//////////////////
          
          

     

    		     

    Neg Ctrl

    • Dummy DNA
    • With Lipofectamine

    Neg Ctrl

  • Dummy DNA
    • With Lipofectamine
    • MAV1212-hEF1a-eBFP2
    • 5xUAS/4xTAL14bs-eYFP
    • hEF1a-Gal4VP16
    • MAV1212-hEF1a-eBFP2
    • 5xUAS/4xTAL14bs-eYFP
    • hEF1a-Gal4VP16
    		//////////////////

    Neg/Neg Ctrl

    • No DNA
    • No Lipofectamin
    • MAV1212-hEF1a-eBFP2
    • 5xUAS/4xTAL14bs-eYFP
    • hEF1a-Gal4VP16
    • hEF1a-TAL14
    • MAV1212-hEF1a-eBFP2
    • 5xUAS/4xTAL14bs-eYFP
    • hEF1a-Gal4VP16
    • hEF1a-TAL14
    		//////

     

    		////////////Neg/Neg Ctrl
    • No DNA
    • No Lipofectamine
    		  

    RESULT: Brief description of experimental outcome (mean transfection efficiency and implications). Link to results page with data analysis.

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    Treatment

     
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