10:00 A – 7:00 P, 9:00 P – 11:55 P
Lab Meeting
- post-doc visit, see notes.
STORM
- Finish O/N imaging of R2bxcS7
- Imaging R2bxcF. high background, lots of blinking = probably cy5 primary + cy5 secondary
- Imaging O/N R2bxcB. low background, very strong signal, high contrast, probably cy5 secondary, unlabeled primary. Not too strongly 405 responsive, though more so perhaps than the last one?
Project 2
see notes
chromatin libraries
- moved to more stringent parameters for hoxD library
- rerunning against newly constructed mouse BLAST database on Odyssey server O/N
Probe making
- RNA cleanup using zymo kit of in vitro transcribed, cy5 labeled RNA probes
- P1 is ~650 ng/uL, RC is ~40 ng/uL. Dye color of solution looks like it.
cell staining
- test probes with previously fixed and prepped Kc167 cells
- En-Primary DNA label (2:20 uL dilution)
- RNA-labeled En-P1 (2:20 uL dilution)
- RNA-labeled En-RC (6:20 uL dilution)
- RNA-labeled En-RC (6:20 uL dilution) + cy5-secondary (1:20 uL dilution) + new-405 (1:20 uL dilution)
- let rubber cement set at 47C.
- 3 min incubation on downstairs heat block at 85C (should be ~78 on glass according to earlier measurments).