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
- AD blood cells from PrecisionMed? Talk to Brian?
- Meet with Kyle about designing siRNA (email sent)
- Make siRNA sequences in geneious
- Send order to Brian
- Picture of how low sensors come together
- Keep updating the parts tree (include experiments)
miRNA Sensors (Theory)
- Determine which pairs of target sites we want
- Find sequences (hopefully under 200 bp)
- Send ultramer order to Brian
- Learn how to acquire MATLAB from Brian
- Find natural concentrations of relevant miRNAs.
miRNA Sensors (Lab Work)
- Golden Gate
- Gather miRNA target sites from Jeremy
- Nanodrop for concentrations ( ? )
- Make all 6 single low sensors (in conjunction with antibody group)
- Gather miRNA target sites from Jeremy
- Create new MAV vector?
- Miniprep eBFP2
- Pick colonies from mKate2 and L7ae
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 Both LR's failed (mKate2 and L7ae)