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)
- More descriptive names
- Include other plasmid
miRNA Sensors (Theory)
- Alternative backbones? Check iGEM 2013
- Plan experiment to test Kturn (Just Kturn and L7ae)
- Re-make double input low sensor as two ultramers (use the Q3 site)
- 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)
- Pick colonies & grow in medium (never just pick one colony)Midi prep
- EF
- GH
- pEXPR: Hef1a-eBFP2
- Mutant Hef1a-2xKturn-eGFP
- Wild Type Hef1a-2xKturn-eGFP
- Mini prep
- pENTR: L7ae
- GH
- Mini prep
- EF
- pEXPR: Hef1a-eBFP2
- Mutant Hef1a-2xKturn-eGFP
- Wild Type Hef1a-2xKturn-eGFP
- LR
- MAV1212-Hef1a-L7ae
- BP
- P4-donor-P1R on Brian's bench
- TAL14-mKate pEST in Brian's fridge
- Golden Gate
- EF
- GH
- Transformation
- BP reaction
- TAL14 mKate
- MAV1212-mRFP
NOTES
- -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 We should sequence the low sensors when they're done