SAMPLE

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

 

Attempt #1

GOAL: Test the strength of L7ae

  • 7/14/14: Seed cells
  • 7/15/14: Transfect MAV1212-hEF1a-eBFP2 (1084.0 ng/uL), Mutant hEF1a-2xKturn-eGFP (551.4 ng/uL) and Wild Type hEF1a-2xKturn-eGFP (287.0 ng/uL) into HEK cells
  • 7/17/14 (2 pm): Flow Cytometer

 

  • DNA Dilution: 5 uL MAV1212-hEF1a-eBFP2, 10 uL Mutant Hef1a-2xKturn-eGFP, 20 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
  • Wild Type Hef1a-2xKturn-eGFP
  • MAV1212-hEF1a-eBFP2

(GM + B + L)2

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

(GWT)1

  • Wild Type Hef1a-2xKturn-eGFP

(GWT)2

  • Wild Type Hef1a-2xKturn-eGFP


(GWT + B)1

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

(GWT + B)2

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

(GWT + B + L)1

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

(GWT + B + L)2

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

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.


Attempt #2

  • 7/16/14: Seed cells
  • 7/17/14: Transfect MAV1212-hEF1a-eBFP2 (1084.0 ng/uL), Mutant hEF1a-2xKturn-eGFP (551.4 ng/uL) and Wild Type hEF1a-2xKturn-eGFP (287.0 ng/uL), MAV1212-hEF1a-L7ae (1686.4 ng/uL) into HEK cells
  • 7/19/14 (2 pm): Flow Cytometer

 

  • DNA Dilution: 5 uL MAV1212-hEF1a-eBFP2, 10 uL Mutant Hef1a-2xKturn-eGFP, 20 uL Wild Type Hef1a-2xKturn-eGFP, 3 uL MAV1212-hEF1a-L7ae

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


(GWT + B)1

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

(GWT + B)2

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

(GWT + B + L)1

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

(GWT + 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

  • Day 1: Seed cells
  • Day 2: Transfect MAV1212-hEF1a-eBFP2 (1084.0 ng/uL), UAS-GAL4-TAL14-eYFP (___ ng/uL), hEF1a-GAL4VP16 (___ ng/uL), hEF1a-GAL4VP16 (___ ng/uL) and hEF1a-GAL4VP16 (___ ng/uL) into HEK cells
  • Day 3: Flow Cytometer

 

  • DNA Dilution: 5 uL MAV1212-hEF1a-eBFP2, ___ uL UAS-GAL4-TAL14-eYFP, ___ uL hEF1a-GAL4VP16, ___ uL hEF1a-TAL14, ___ UAS-GAL4-TAL14-mKate


24-WELL PLATE TRANSFECTION MAP:

(B)1

  • MAV1212-hEF1a-eBFP2

(B)2

  • MAV1212-hEF1a-eBFP2
////////////

//////

 

Neg Ctrl

  • Dummy DNA
  • With Lipofectamine

(B + Y)1

  • MAV1212-hEF1a-eBFP2
  • 5xUAS/4xTAL14bs-eYFP

(B + Y)2

  • MAV1212-hEF1a-eBFP2
  • 5xUAS/4xTAL14bs-eYFP

(B + R)1

  • MAV1212-hEF1a-eBFP2
  • 5xUAS/4xTAL14bs-mKate

(B + R)2

  • MAV1212-hEF1a-eBFP2
  • 5xUAS/4xTAL14bs-mKate
//////

Neg Ctrl

  • Dummy DNA
  • With Lipofectamine

(B + Y + GAL)1

  • MAV1212-hEF1a-eBFP2
  • 5xUAS/4xTAL14bs-eYFP
  • hEF1a-Gal4VP16

(B + Y + GAL)2

  • MAV1212-hEF1a-eBFP2
  • 5xUAS/4xTAL14bs-eYFP
  • hEF1a-Gal4VP16

(B + R + GAL)1

  • MAV1212-hEF1a-eBFP2
  • 5xUAS/4xTAL14bs-mKate
  • hEF1a-Gal4VP16

(B + R + GAL)2

  • MAV1212-hEF1a-eBFP2
  • 5xUAS/4xTAL14bs-mKate
  • hEF1a-Gal4VP16
//////

Neg/Neg Ctrl

  • No DNA
  • No Lipofectamin

(B + Y + GAL)1

  • MAV1212-hEF1a-eBFP2
  • 5xUAS/4xTAL14bs-eYFP
  • hEF1a-Gal4VP16
  • hEF1a-TAL14

(B + Y + GAL)2

  • MAV1212-hEF1a-eBFP2
  • 5xUAS/4xTAL14bs-eYFP
  • hEF1a-Gal4VP16
  • hEF1a-TAL14

(B + R + GAL)1

  • MAV1212-hEF1a-eBFP2
  • 5xUAS/4xTAL14bs-mKate
  • hEF1a-Gal4VP16
  • hEF1a-TAL14

(B + R + GAL)2

  • MAV1212-hEF1a-eBFP2
  • 5xUAS/4xTAL14bs-mKate
  • 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.

Treatment