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)
- Update experiments page
- Obtain plasmid maps
- Email Jin about TAL14 and miRNA target sites
- Talk to Lyla/Alexa about transfecting later this week
- Look for plasmids
- pEXPR: UAS-Gal4-TAL14-eYFP
- pEXPR: Gal4VP16
- pEXPR: Hef1a-TAL14
- pEXPR: Hef1a-2x Kturn-eYFP
- 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)
- Transformations
- Low Sensor Golden Gates
- pEXPR: Hef1a-eBFP2 (186 ng/uL)
- pEXPR: Hef1a-L7ae (96 ng/uL)
- Pick red colonies and grow in medium!
- Find concentrations of eBFP2 and L7ae
NOTES
Nelson doesn't know where it is, but he needs it as well, so he's on the hunt
If we're ready, we can transfect very early next week
- 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