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Experiment #1

GOAL: Test promoters in the absence/presence strength of repressorsL7ae

3 incomplete Day 1: Seed cells 22 incomplete Day 2: Transfect MAV1212-hEF1a-eBFP2 (___ ng/uL - transfection marker) into HEK cells 0 incomplete Day 3: Flow Cytometer

 

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


24-WELL PLATE TRANSFECTION MAP:

 hEFa1-L7ae

-ve control

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

    (B)1

    • MAV1212-hEF1a-eBFP2

    (B)2

    • MAV1212-hEF1a-eBFP2
    		   

    -ve control

    • Dummy DNA
    • With Lipofectamine


    -ve control

    • Dummy DNA
    • With Lipofectamine


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

    (GM)1

    • Mutant Hef1a-2xKturn-eGFP

    (GM)2

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

    (GWT)1

    • Wild Type Hef1a-2xKturn-eGFP

    (GWT)2

    • Wild Type Hef1a-2xKturn-eGFP


    • Wild Type Hef1a-2xKturn-eGFP
    • MAV1212-hEF1a-eBFP2
    • Wild Type Hef1a-2xKturn-eGFP
    • MAV1212-hEF1a-eBFP2
    • Wild Type Hef1a-2xKturn-eGFP
    • MAV1212-hEF1a-eBFP2
    • hEF1ahEFa1-L7ae

    -ve control

    • Dummy DNA
    • With Lipofectamine
    		-ve/-ve control
    • No DNA
    • No Lipofectamin
    • Wild Type Hef1a-2xKturn-eGFP
    • MAV1212-hEF1a-eBFP2
    • hEF1a-L7ae
          


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

    Experiment #2

    GOAL: Test promoters in the presence of repressors

    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) 7 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.

    ...