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Experiment #1

Attempt #1

GOAL: Test the strength of L7ae

3 complete 7/14/14: Seed cells 22 complete 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 0 incomplete 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

2 complete 7/16/14: Seed cells 1 incomplete 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 1 7 incomplete 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 analysi

Experiment #2

GOAL: Test the strength of TAL14

14 incomplete Day 1: Seed cells 15 incomplete Day 2: Transfect MAV1212-hEF1a-eBFP2 (1084.0 ng/uL), UAS-GAL4-TAL14-eYFPmKate (___ ng/uL), hEF1a-GAL4VP16 (___ ng/uL), hEF1a-GAL4VP16TAL14 (___ ng/uL) and hEF1a-GAL4VP16and UAS-TAL14-eYFP (___ ng/uL) into HEK cells 16 incomplete Day 3: Flow Cytometer

 

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


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/4xTAL14bsUAS-TAL14-eYFP

(B + Y)2

  • MAV1212-hEF1a-eBFP2
  • 5xUAS/4xTAL14bsUAS-TAL14-eYFP

(B + R)1

  • MAV1212-hEF1a-eBFP2
  • 5xUAS/4xTAL14bsUAS-TAL14-mKate

(B + R)2

  • MAV1212-hEF1a-eBFP2
  • 5xUAS/4xTAL14bsUAS-TAL14-mKate
//////

Neg Ctrl

  • Dummy DNA
  • With Lipofectamine

(B + Y + GAL)1

  • MAV1212-hEF1a-eBFP2
  • 5xUAS/4xTAL14bsUAS-TAL14-eYFP
  • hEF1a-Gal4VP16

(B + Y + GAL)2

  • MAV1212-hEF1a-eBFP2
  • 5xUAS/4xTAL14bsUAS-TAL14-eYFP
  • hEF1a-Gal4VP16

(B + R + GAL)1

  • MAV1212-hEF1a-eBFP2
  • 5xUAS/4xTAL14bsUAS-TAL14-mKate
  • hEF1a-Gal4VP16

(B + R + GAL)2

  • MAV1212-hEF1a-eBFP2
  • 5xUAS/4xTAL14bsUAS-TAL14-mKate
  • hEF1a-Gal4VP16
//////

Neg/Neg Ctrl

  • No DNA
  • No Lipofectamin

(B + Y + GAL)1

  • MAV1212-hEF1a-eBFP2
  • 5xUAS/4xTAL14bsUAS-TAL14-eYFP
  • hEF1a-Gal4VP16
  • hEF1a-TAL14

(B + Y + GAL)2

  • MAV1212-hEF1a-eBFP2
  • 5xUAS/4xTAL14bsUAS-TAL14-eYFP
  • hEF1a-Gal4VP16
  • hEF1a-TAL14

(B + R + GAL)1

  • MAV1212-hEF1a-eBFP2
  • 5xUAS/4xTAL14bsUAS-TAL14-mKate
  • hEF1a-Gal4VP16
  • hEF1a-TAL14

(B + R + GAL)2

  • MAV1212-hEF1a-eBFP2
  • 5xUAS/4xTAL14bsUAS-TAL14-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.

...