10:00 am – 9:00 pm
STORM
L2 F06F07
- finish hot-washes
- add beads, make STORM buffer
- Imaging F06-p3-7 F07-p2-6 double color
- staining in both channels bright and clear
- colocalization looks decent
New buffer for multicolor STORM
- 2.5 uL MEA
- 1.5 uL BME
- 5 uL COT
- 10 uL GLOX
Writing
- writing up formal comments on manuscript for colleagues
- # need to write abstract of DR retreat!
New Probe making
regions
- internal black domains: A01-A10 + E01-E10
- some BXC small pieces: A11-B12 + E11-F12
- 24 new probe regions in all (plus let’s do some in 2 color).
per well
- 25 uL 2x Phusion master hot start
- 2.5 uL Eva Green
- 5 uL library
- 12.5 uL ddH2O
- 2.5 uL Fwd Primer
- 2.5 uL Rev Primer
27 x master mix
- 675 uL Phusion hot start master mix
- 67.5 uL Eva Green
- 135 uL library
- 337.5 uL ddH2O
Cleanup
- ran cleanup using Zymo 96 well plate.
- much easier than individual spin columns