9:30 A – 7:30 P
- Finish comments on counting comparison for ML
- # Finish revisions Write to MERCK about
Probe Making
- Precipitate and resuspend RNA-FISH DNP probes for tou, inv, en, Antp and Dfd.
- Make new RNA-Hybe soln to resuspend probes in
- start synthesis of en DNA OligoPaint probes when (if?) primers arrive
- 25 nmole of DNA. in 125 uL = 200 uM. Need 50 uL per 100x set of reactions, should probably have ordered in the 250 nmole scale…
Coding
- working on STORMfinder
- basic dot-finding paramter manipulation for InsightM working.
- Starting on writing parameter GUI for DaoSTORM.
- Matlab froze.
- Reboot, resume coding
Update Protocols page
Cell Culture
- split cells for all three cell lines.
- Plate cells in new 8 well chambered slide to attempt centromere FISH again using my previous protocol (which unfortunately needs a larger volume of probe, but better to burn through probe than not know if it’s good or not)
- Chose two wells, repeating at 5:150 uL dilution + previous probe recovered from slide diluted down to 150.
Fly work
- water all 50 back-crosses from recombineering screen (first 25 lines). Lines struggling but we’ll see what happens — hopefully we can still score a good fraction of them.
- MTD screening cross of shRNA definetely struggling (1 – 2 pupae per vial, not looking good for making a decent F2 generation).
- All potential virgin vials look clean. Moved to 18C. Should use the nice collection of MTDs to repeat the crosses that failed when started at 29C…