eSAMPLE

Sample Experiment #1 (Template)

  • Day 1: Seed cells
  • Day 2: Transfect Promoter:Gene1 (___ ng/uL - transfection marker), Promoter:Gene2 (___ ng/uL) and Promoter:Gene3 (___ ng/uL) into HEK cells
  • Day 3: Add DOX (if necessary)
  • Day 4 (time): Flow Cytometer

  • DNA Dilution: __uL Promoter:Gene1; __uL Promoter:Gene2; __uL Promoter:Gene3
  • (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:

 

+ve control
  • Promoter:Gene1
+ve control
  • Promoter:Gene1
-ve control
  • dummy DNA
  • with Lipofectamine
-ve control
  • dummy DNA
  • with Lipofectamine
-ve/-ve control
  • no DNA
  • no Lipofectamine
-ve/-ve control
  • no DNA
  • no Lipofectamine
///EMPTY WELL/////////////////////////////////
////////////////////////////////////
(Eg)1
  • __uM DOX
  • Promoter:Gene1
  • Promoter:Gene2
  • Promoter:Gene3
(Eg)2
  • __uM DOX
  • Promoter:Gene1
  • Promoter:Gene2
  • Promoter:Gene3
(Eg)3
  • __uM DOX
  • Promoter:Gene1
  • Promoter:Gene2
  • Promoter:Gene3
(Eg)4
  • __uM DOX
  • Promoter:Gene1
  • Promoter:Gene2
  • Promoter:Gene3
(Eg)5
  • __uM DOX
  • Promoter:Gene1
  • Promoter:Gene2
  • Promoter:Gene3
(Eg)6
  • __uM DOX
  • Promoter:Gene1
  • Promoter:Gene2
  • Promoter:Gene3

 

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

Sample Experiment #2

(This was Practice Experiment #2)

  • 7/3: Seed cells
  • 7/4: Transfect hEF1a:eYFP (472 ng/uL) and hEF1a:tagBFP (1176 ng/uL) into HEK cells
  • 7/6 (1pm): Flow Cytometer

 

  • DNA Dilution: 10uL hEF1a:eYFP; 5uL hEF1a:tagBFP
  • (Y+B)x: add 1:1 (25uL hEF1a:eYFP/tagBFP-lipid complex)


Y1
  • hEF1a:eYFP
(Y+B)1
  • hEF1a:eYFP
  • hEF1a:tagBFP
//////////////////-ve control
  • dummy DNA
  • with Lipofectamine
Y2
  • hEF1a:eYFP
(Y+B)2
  • hEF1a:eYFP
  • hEF1a:tagBFP
//////////////////-ve control
  • dummy DNA
  • with Lipofectamine
B1
  • hEF1a:tagBFP
(Y+B)3
  • hEF1a:eYFP
  • hEF1a:tagBFP
//////////////////-ve/-ve control
  • no DNA
  • no Lipofectamine
B2
  • hEF1a:tagBFP
(Y+B)4
  • hEF1a:eYFP
  • hEF1a:tagBFP
//////////////////-ve/-ve control
  • no DNA
  • no Lipofectamine


RESULT: Extremely low transfection efficiency (<10%). Repeat to improve co-transfection results.

 

Antibody Detection

 

Receptor Detection

 

miRNA Detection

Experiment #1

GOAL: Test the strength of L7ae

  • 7/14/14: Seed cells
  • 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
  • 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

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 L7ae

  • Day 1: Seed cells
  • Day 2: Transfect Promoter:Gene1 (___ ng/uL - transfection marker), Promoter:Gene2 (___ ng/uL) and Promoter:Gene3 (___ ng/uL) into HEK cells
  • Day 3: Add DOX (if necessary)
  • 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.

Treatment