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| TASK | WhoDunnit | Observed |
|---|
| Split cells | AG, CR, LA, SS | AW |
| Seed plates | AG, CR, LA, SS | AW |
| Transfection | AG, LA | AC, SS |
| FACs Prep | LA | CR |
| Imaging (FloCyto) | BT | AG, AW, CR, JX, LA, RR |
Practice Transfections:
CONTROLS:
- -ve/-ve: no actions performed on cells
- -ve: perform protocol, excluding DNA, or with dummy DNA
- +ve: tri-color control (perform experiments with YFP, BFP, mKATE separately, and one with all 3: allows for comparison of color/intensity when imaging)
PRACTICE #1: Fluorescent protein under constitutive promoter
- 6/23: Seed cells
- 6/24: Transfect hEF1a:eYFP into HEK cells
- 6/26 (4.30pm): Flow Cytometer
- Determine optimal Lipofectamine LTX volume (24 well)
- Determine transfection efficiency (quantitatively)
PRACTICE #2: Fluorescent protein under inducible promoter
- 7/2: Seed cells
- 7/3: Transfect TRE:mKate and TRE:eYFP into HEK cells
- 7/4: Add DOX
- 7/5 (__pm): Flow Cytometer
PRACTICE #3: Co-transfection (2 fluorescent proteins under constitutive promoter)
- 7/_: Seed cells
- 7/_: Transfect hEF1a:____ and hEF1a:____ into HEK cells
- 7/_ (__pm): Flow Cytometer
PRACTICE #4: Co-transfection (2 fluorescent proteins under constitutive & inducible promoters)
- 7/_: Seed cells
- 7/_: Transfect hEF1a:____ and TRE:____ into HEK cells
- 7/_: Add DOX
- 7/_ (__pm): Flow Cytometer