10:00 A – 6:45 P, 8:30 P – 11:30 P
Ph project
- washing out primary antibodies
- stain with new secondary antibodies.
- wash out secondaries
- Prep samples for STORM
STORM
- O/N imaging of Ph-wt Ph-cy7 + Pc-647
- 750 mosaic acquired at 700 mA 750 laser
- 1600 mA 750, 50% AOTF 180 mW 647
- Bead movie acquisition: stage froze at second position, need to re-aquire beads.
- dye switching and preliminary movies look good.
chromatin
- discuss reverse transcriptase option
Review writing
- Pedraza and Paulsson observe a .3 to .2 transition as an effect of ‘gestation’ by ensuring that the firing rates are equal (N transitions in the N-step model all are N times as fast as the 1 step model, so the average activation rate is the same) keeping mRNA burst size small (burst size 1, keeps the non-reducible noise low) by examining a large protein burst regime (~8 mRNA, 500-3500 proteins. Since coefficient of variation scales with burst size, a small reduction in mRNA CoV still results in a large reduction in protein CoV.
- These bursts sizes are probably reasonable for some genes, which suggests that there must be even more pressure on slight variations in mRNA levels.
- working on rewriting gestation section and sketching figure