Sample Experiment #1 (Template): 4 complete 7/_ incomplete Day 1: Seed cells 5 complete 7/_ incomplete Day 2: Transfect Promoter:Gene1 (___ ng/uL - transfection marker), Promoter:Gene2 (___ ng/uL) and Promoter:Gene3 (___ ng/uL) into HEK cells 6 complete 7/_ (_pm 8 incomplete Day 3: Add DOX (if necessary) 6 incomplete Day 4 (time): Flow Cytometer
- DNA Dilution: __uL Promoter:Gene1; __uL Promoter:Gene2; __uL Promoter:Gene3
- (Eg)x: add : (25uL _:_:_
- __uL Promoter:Gene1-lipid complex
)- __uL Promoter:Gene2-lipid complex
- __uL Promoter:Gene3-lipid complex
24-WELL PLATE TRANSFECTION MAP: 36 complete 7| +ve control | +ve control | -ve control- dummy DNA
- with Lipofectamine
|
| Y1 | (Y+B)1 | ////// | ////// | ////// | -ve control- dummy DNA
- with Lipofectamine
| Y2 | (Y+B)2 | //////-ve/-ve control | -ve/-ve control | | ///EMPTY WELL/// | ////// | -ve controldummy DNAwith Lipofectamine | B1 | (Y+B)3 | ////// | ////// | ////// | -ve/ | -ve controlB2 | hEF1a:tagBFP(Y+B)4 | ////// | | ////// | ////// | -ve/ | -ve control
|---|
DNA Dilution: 10uL hEF1a:eYFP; 5uL hEF1a:tagBFP(Y+B)x: add 1:1 (25uL hEF1a:eYFPtagBFP-lipid complex) Y1 | (Y+B)1hEF1a:eYFPhEF1a:tagBFP-ve control- dummy DNA
- with Lipofectamine
| | Y2 | (Y+B)2 | ////// | ////// | ////// | -ve control- dummy DNA
- with Lipofectamine
| | B1 | (Y+B)3 | ////// | ////// | ////// | -ve/-ve control | (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
|
B2 | (Y+B)4 | ////// | ////// | ////// | -ve/-ve controlno DNAno Lipofectamine |