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)
- Anaylze Low Sensor miR-144 sequencing resultsRe-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)
- Restriction DigestMAV1212Midi-hEF1a-L7ae / that other shit in the fridge
- Golden Gate
- High Sensor miR-30d (I13 & J13)
- High Sensor miR-146a (I5 & J5)
- High Sensor let-7f (1.13 & 1.14)
- High Sensor miR-125b (A3 & B3)
- Low Sensor miR-144 (E5 & F5)
- Low Sensor miR-181c (G7 & H7)
- Low Sensor miR-144-miR-181c
- Transform
- pEXPR: MAV1212-TRE-L7ae (LR)
- pEXPR: MAV1212-hEF1a-GAL4VP16 (LR)
- pEXPR: MAV1212-TRE-GAL4VP16 (LR)
- Re-suspend Low Sensor miR-144-181c oligos
- Anneal Low Sensor miR-44-181c oligos
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 Low Sensor miR-144 did not pass quality control for sequencing