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)
- 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)
- Midi-prep
- pEXPR: MAV1212-hEF1a-TAL14
- Transform
- pEXPR: MAV1212-TRE-L7ae (LR from last night)
- Proteinase K
- MAV1212-hEF1a-TAL14
- Restriction Digest and run gel
- MAV1212-hEF1a-TAL14
- MAV1212-hEF1a-TAL14
- LR
- MAV1212-hEF1a-TAL21
- Mini prep
- Low Sensor miR-144
- Low Sensor miR-181c
- High Sensor miR-30d
- High Sensor miR-125b
- High Sensor miR-146a
- High Sensor let-7f
- Send for sequencing
- Low Sensor miR-144
- Low Sensor miR-181c
- High Sensor miR-30d
- High Sensor miR-125b
- High Sensor miR-146a
- High Sensor let-7f
- Pick colonies
- Low Sensor miR-144
- Low Sensor miR-181c
- High Sensor miR-30d
- High Sensor miR-125b
- High Sensor miR-146a
- High Sensor let-7f
- pEXPR: MAV1212-hEF1a-GAL4VP16
- pEXPR: MAV1212-TRE-GAL4VP16
- Seed plates
- L7ae experiment (Take 3)
NOTES
- 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