9:45 A – 8:30 P
- write to ML about bistability
- write to Bender about Hox model. — apparently email contact is not recommended. Try phone tomorrow.
- write summary of Francis suggestions
- remove medium from S2 cells, add fresh medium. Reduce total volume to 50 mL.
- write to Eric about MBNL
- send in PO for latest fly order (PcG mutants, marked balancer 3, hs-en).
Meeting with Fred and Brian
- got 20kb Fab7 BAC probe with A488 from Fred (100 ng of probe. Dilute in 30 uL of DFISH buffer with embryos)
- Got probe template and labeled primer set from Brian and Nick. 50 kb region
- Brian will send browser for designing probes (basically has pre-tiled Drosophila genome with ideal probes (repeat screened, TM optimized etc) as a BED file. Simply intersect ROI BED file with this data set to get probe set.
- check sample. Focus lossed O/N. Reset focus. should keep an eye on this, will probably continue to get lost. Monitor with remote desktop.
- Switching still looks similar to yesterday, Pyranose system working so far.
- switching looking a little sparse, may be cell type.
- lost focus again (<11:30A), system very sensitive 🙁 — maybe a double spot issue.
- Getting substantially fewer localizations from 750 channel. May be buffer issue, may be section issue? Will let scope try out a few more sections before re-etching.
- substantial number of localizatios in 647 channel not turning off properly. really should expand linkage and throw out long-ON dots more. Better filtering needed and should not be too hard.
- 750 density becoming pretty crappy. 647 looks qualitatively good still but total localization may have dropped substantially.