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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- dummy DNA
- with Lipofectamine
| -ve control- dummy DNA
- with Lipofectamine
| -ve/-ve control | -ve/-ve control |
| ///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 | (Y+B)1 | ////// | ////// | ////// | -ve control- dummy DNA
- with Lipofectamine
|
| Y2 | (Y+B)2 | ////// | ////// | ////// | -ve control- dummy DNA
- with Lipofectamine
|
| 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
Treatment