Long Term Goals
miRNA Profile
- Read literature
- Choose profile
Building the Cell Detector Circuit
- Build Choose profile
- Verify sensors
- Create mathematical model
- Make test bed
- Controlled miRNA expression
- Constructed miRNA sensors
Mini experiments
- Single miRNA detector circuit
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)
- Figure out where the Golden Gates went
- Create protocol for siRNA transfection
- Meet with Jin at 3
- 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)
- Transformations
- EF
- GH
- Mutant Hef1a-2xKturn-eGFP
- Wild Type Hef1a-2xKturn-eGFP
- Pick colonies from L7ae and eBFP2
- Miniprep MAV1212
- Digest MAV1212
- Run gel for MAV1212
- Image gel for MAV1212
NOTES
- Midi-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 Jin gave us ideas to optimize our high sensors and experiments