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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 _:_:_
- __uL Promoter:Gene1-lipid complex
- __uL Promoter:Gene2-lipid complex
- __uL Promoter:Gene3-lipid complex
24-WELL PLATE TRANSFECTION MAP:
+ve control
| +ve control
| -ve control
| -ve control
| -ve/-ve control
| -ve/-ve control
|
| ///EMPTY WELL/// | ////// | ////// | ////// | ////// | ////// |
| ////// | ////// | ////// | ////// | ////// | ////// |
|---|---|---|---|---|---|
(Eg)1
| (Eg)2
| (Eg)3
| (Eg)4
| (Eg)5
| (Eg)6
|
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
| (Y+B)1
| ////// | ////// | ////// | -ve control
|
Y2
| (Y+B)2
| ////// | ////// | ////// | -ve control
|
B1
| (Y+B)3
| ////// | ////// | ////// | -ve/-ve control
|
B2
| (Y+B)4
| ////// | ////// | ////// | -ve/-ve control
|
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 (12 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
| (B)2
| ////// | ////// | ////// | ////// |
(GM)1
| (GM)2
| (GM + B)1
| (GM + B)2
| (GM + B + L)1
| (GM + B + L)2
|
(GWT)1
| (GWT)2
| (GWT + B)1
| (GWT + B)2
| (GWT + B + L)1
| (GWT + B + L)2
|
Neg Ctrl
| Neg Ctrl
| Neg/Neg Ctrl
| Neg/Neg Ctrl
| ////// | ////// |
RESULT: Transfection efficiency ~35%. FACS data is available here
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 (10 am): 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
| (B)2
| ////// | ////// | ////// | ////// |
(GM)1
| (GM)2
| (GM + B)1
| (GM + B)2
| (GM + B + L)1
| (GM + B + L)2
|
(GWT)1
| (GWT)2
| (GWT + B)1
| (GWT + B)2
| (GWT + B + L)1
| (GWT + B + L)2
|
Neg Ctrl
| Neg Ctrl
| Neg/Neg Ctrl
| Neg/Neg Ctrl
| ////// | ////// |
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
- Day 1: Seed cells
- Day 2: Transfect MAV1212-hEF1a-eBFP2 (1084.0 ng/uL), UAS-TAL14-mKate (___ ng/uL), hEF1a-GAL4VP16 (___ ng/uL), hEF1a-TAL14 (___ ng/uL) and UAS-TAL14-eYFP (___ ng/uL) into HEK cells
- Day 4: Flow Cytometer
- DNA Dilution: 5 uL MAV1212-hEF1a-eBFP2, ___ uL UAS-TAL14-mKate, ___ uL hEF1a-GAL4VP16, ___ uL hEF1a-TAL14, ___ UAS-TAL14-eYFP
24-WELL PLATE TRANSFECTION MAP:
(B)1
| (B)2
| ////// | ////// | //////
| Neg Ctrl
|
(B + Y)1
| (B + Y)2
| (B + R)1
| (B + R)2
| ////// | Neg Ctrl
|
(B + Y + GAL)1
| (B + Y + GAL)2
| (B + R + GAL)1
| (B + R + GAL)2
| ////// | Neg/Neg Ctrl
|
(B + Y + GAL)1
| (B + Y + GAL)2
| (B + R + GAL)1
| (B + R + GAL)2
| ////// | Neg/Neg Ctrl
|
RESULT: Brief description of experimental outcome (mean transfection efficiency and implications). Link to results page with data analysis.
Experiment #3
GOAL: Test the sensors in absence of siRNA
- Day 1: Seed cells
- Day 2: Transfect Low Sensor miR-144 (___ ng/uL), Low Sensor miR-181c (___ ng/uL), High Sensor miR-30d (___ ng/uL), High Sensor miR-146a (___ ng/uL), High Sensor 125b (___ ng/uL) and High Sensor let-7f (___ ng/uL) into HEK cells
- Day 4: (2 pm): Flow Cytometer
- DNA Dilution: ___ uL Low Sensor miR-144, ___ uL Low Sensor miR-181c, ___ uL High Sensor miR-30d, ___ uL High Sensor miR-146a, ___ uL High Sensor 125b, ___ uL High Sensor let-7f
24-WELL PLATE TRANSFECTION MAP:
144
|
|
|
|
|
|
|
|
|
|
|
|
////// | ////// | ////// | ////// | ////// | ////// |
Neg Ctrl
| Neg Ctrl
| Neg/Neg Ctrl
| Neg/Neg Ctrl
|
////// |
////// |
RESULT: Brief description of experimental outcome (mean transfection efficiency and implications). Link to results page with data analysis.
Experiment #4
GOAL: Test the sensors in the presence of siRNA
- Day 1: Seed cells
- Day 2: Transfect Low Sensor miR-144 (___ ng/uL), Low Sensor miR-181c (___ ng/uL), High Sensor miR-30d (___ ng/uL), High Sensor miR-146a (___ ng/uL), High Sensor 125b (___ ng/uL), High Sensor let-7f (___ ng/uL), Mature 144 (___ ng/uL), Mature 181c (___ ng/uL), Mature 30d (___ ng/uL), Mature 146a (___ ng/uL), Mature 125b (___ ng/uL) and Mature let-7f (___ ng/uL) into HEK cells
- Day 4: (2 pm): Flow Cytometer
- DNA Dilution: ___ uL Low Sensor miR-144, ___ uL Low Sensor miR-181c, ___ uL High Sensor miR-30d, ___ uL High Sensor miR-146a, ___ uL High Sensor 125b, ___ uL High Sensor let-7f , ___ uL Mature 144, ___ uL Mature 181c, ___ uL Mature 30d, ___ uL Mature 146a, ___ uL Mature 125b and ___ uL Mature let-7f
24-WELL PLATE TRANSFECTION MAP:
144
|
|
|
|
|
|
|
|
|
|
|
|
////// | ////// | ////// | ////// | ////// | ////// |
Neg Ctrl
| Neg Ctrl
| Neg/Neg Ctrl
| Neg/Neg Ctrl
|
////// |
////// |
RESULT: Brief description of experimental outcome (mean transfection efficiency and implications). Link to results page with data analysis.
Treatment
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