10:00 A – 12:30 A
cell staining
- heating wash buffers to 47C
- imaged 20 bp single probe cells (5 cells).
- imaged 40 bp array (with 0 gain, oops). (6 cells)
- imaged 40 bp array for restain (7 cells)
- Doory imaging rest of day.
- relabeling 40bp array with Cy3 at 37C. Started at 1:30P, run through tomorrow.
- washout 60C 1:30P – 8:00P. image washout
- restain (210 uL hybe buffer, 60 uL 50% dextran, 30 uL probe)
dye test:
- stain 40 bp long cy5,cy7 and cy3,cy5 probes (2 of each).
- 1x PBS rinse,
- PBT 5% FA fix,
- 2x PBS rinse
- 20 min -20C ethanol permiabilization
- PBS rinse,
- 2x SSC rinse
- prehybe 35 min
- hybe O/N 37C: (30 uL probe, 75 uL hybe buffer, 30 uL 50% dextran) oops. wanted 15 75 30 or 30 110 60
- forgot sodium borohydride!!
embryo staining
- heating wash chamber and 2x SSCT buffer to 60C
- transfer to SSC
- move to 4C at 11:30 PM (not going to be able to image today)
Presentation Prep
Slides for Meeting with XZ today
- text outline: recap, summary of progress, goals for next 2 weeks
Slides for SDB Presentation
- checking data on Alistair3 for Pc-H3K27 Psc-H3K27 association
- images of interocolated and non-intercolated overlapping DNA
Slides for Project 2 team meeting today
Slides for Ting Wu group meeting
Meetings
- team-project meeting with XZ (see notes)
- individual meeting with XZ (see notes)
Ph
- Ajaz brought samples. Need to image
- Brian brought samples. Need to image