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)
- Alternative backbones (check iGEM 2013)
- Obtain pEXPR: hEF1a-GAL4VP16 from Brian
- Fill out the Genewiz Order Form
- Prepare transfection well plate maps
- So where are the siRNA though...
- Why didn't the GH golden gate work on try #3?
- Change name of high sensors in tree
- 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)
- Prepare low sensors for sequencing (protocol on the wall of the lab)
- Pick colonies
- Hef1a-TAL14 (LR)
- pEXPR: TAL14-mKate
- Midiprep
- pEXPR: MAV1212-Hef11a-eBFP2
- Mutant Hef1a-2xKturn-eGFP
- Wild Type Hef1a-2xKturn-eGFP
- Low Sensor miR-144
- Hef1a-L7ae (LR)
- Miniprep
- MAV1212-mRFP
- L1_TALER14_L2
- pTAL14-mKate-pEST
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