Long Term Goals
Building the Cell Detector Circuit
- Choose profile
- Verify sensors
- Create mathematical model
Experiments
- L7ae repression
- TAL14 repression
- TAL21 repression
- Single input sensors
- Multiple input sensors
Alzheimer's Model
- Agree on a team wide, full Alzheimer's model
Today's Agenda
How To Please Ron/Brain
- Keep updating the parts tree (include experiments)
miRNA Sensors (Theory)
- Get TAL21 promoter from Brian
- Plan experiments
- DOX induction for L7ae repression (look here)
- Tuning TAL14 repression
- Learn about gating and FACS analysis
- Re-analyze all of our data in MATLAB (Gary)
- Learn how to acquire MATLAB from Brian
- Find natural concentrations of relevant miRNAs.
miRNA Sensors (Lab Work)
- Midi prep
- pEXPR: MAV1212-TRE-L7ae
- Mini prep
- MAV1212-hEF1a-TAL21
- MAV1212-hEF1a-TAL14
- Low Sensor miR-144-181c
- Sequence verify
- Low Sensor miR-144-181c
- Digest and run gel
- MAV1212-hEF1a-TAL21
- MAV1212-hEF1a-TAL14
- Golden Gate (if digest works)
- All the TAL21 high sensors
- Transform
- All the TAL14 high sensors
- And a plasmid from BCR
- -prep hEF1a:rtTA and hEF1a: Gal4VP16 2A rtTA3
- Mini-prep MAV1212 hEF1A Tal14
- Digest and Map MAV1212 hEF1A Tal14
- Transform third LR trial if not verified
- Pick/Streak hEF1a: Gal4VP16 2A rtTA3
NOTES:
hEF1a: Gal4VP16 2A rtTA3 culture did not grow so NOTES