Excerpt From Lab Notebook

This is why it's taking a while to get my paper . . .

3-15-06
Plated pax -/- MEFs from August 2005 in freshly made high-glucose DMEM plus 15% BCS, 1% GPS. Filtered medium .2 mm filter.

Took images on Leica from the following slides:
Pax+ Pkl red (tho marked green) Zyx G (tho…) Zyx 1:250, Pkl 1:50, 1:100
Pax -/- Zyx G, Pkl red, (unstim? On Fn?) from 10-21-05

Focused using 488 laser (zyx) at 15-20% and set gain of red channel so that there was little/no bleedthrough. Then imaged using same settings for green channel and gains, with 594 laser at 100%.

PaxC cells – can see myc staining and Pkl staining but didn’t take pictures. They look like fractals bc of all of the ruffling ruffles coming out of other ruffles.

3-16-05
Cells sparse and I see some of the little blobs as well as some stringy tails on cells. Shit.

Plated another vial from October 2004 in same medium. (The bottle of which I then dropped on the floor.) Two more vials from 10-04 and 6-04 in box 3-18G. (Has it been that long???) Also have N and C mutants and at least one wildtype in that box. A few later vials also in box 1-2B.

Make sure to do 2° Ab alone control for Pkl next time and use TBS-T for washes as per ref.