| TASK | WhoDunnit | Observed |
|---|---|---|
| Split cells | AG, CR, LA, SS | AW |
| Seed plates | AG, CR, LA, SS | AW |
| Transfection | AG, LA | AC, SS |
| Imaging (FloCyto) |
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
- 6/26: Cells seeded
- 6/27: Transfect TRE:mKate and TRE:eYFP into HEK cells
- 6/29 (__pm): Flow Cytometer