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)
- Troubleshoot
- What's the deal with the GAL4VP16?!
- Learn how to acquire MATLAB from Brian
- Find natural concentrations of relevant miRNAs.
miRNA Sensors (Lab Work)
- Pick colonies
- pEXPR: MAV1212-TRE-L7ae (LR from last night)
- MAV1212-hEF1a-GAL4VP16
- MAV1212-TRE-GAL4VP16
- Proteinase K
- MAV1212-hEF1a-TAL21
- MAV1212-hEF1a-TAL14
- Transform
- MAV1212-hEF1a-TAL21
- MAV1212-hEF1a-TAL14
- Miniprep
- All the sensors (round 2)
- Send for sequencing
- All the sensors (round 2)
- Analyze sequencing results
- All the sensors (round 1)
- Transfect
- L7ae Experiment (Take 3)
- Seed
- TAL14 Experiment (Take 1)
- Re-anneal
- miR-144-181c double input target site
- Golden Gate
- Low Sensor miR-144-181c
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 All of the low sensors were verified!