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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 + B)1

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

(GM + B)2

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

(GM + B + L)1

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

(GM + B + L)2

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

-ve controlNeg Ctrl

  • Dummy DNA
  • With Lipofectamine


-ve controlNeg Ctrl

  • Dummy DNA
  • With Lipofectamine


-ve/-ve control

Neg/Neg Ctrl

  • No DNA
  • No Lipofectamin
-ve/-ve control

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 L7ae

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 3: Add DOX (if necessary) 10 incomplete Day 4 (time): Flow Cytometer
  • DNA Dilution: ___ uL Mutant Hef1a-2xKturn-eGFP, ___ uL Wild Type Hef1a-2xKturn-eGFP, ___ uL hEF1a-L7ae
  • (Eg)x: add _:_:_
    1. __uL Promoter:Gene1-lipid complex
    2. __uL Promoter:Gene2-lipid complex
    3. __uL Promoter:Gene3-lipid complex


24-WELL PLATE TRANSFECTION MAP:

GWT

  • Wild Type Hef1a-2xKturn-eGFP

GM

  • Mutant Hef1a-2xKturn-eGFP
 ////////////-ve control
  • Dummy DNA
  • With Lipofectamine

GWT

  • Wild Type Hef1a-2xKturn-eGFP

GM

  • Mutant Hef1a-2xKturn-eGFP
 ////////////-ve control
  • Dummy DNA
  • With Lipofectamine

 


  ////////////-ve/-ve control
  • No DNA
  • No Lipofectamine

 


  ////////////-ve/-ve control
  • No DNA
  • No Lipofectamine

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

...