9:40 am – 11:00 pm
Heat shock experiments
- continue cell prep
- LN2 treatment,
- 5 min 1M HCl bath
- RNase A digestion (30 min at 37C)
- SSCT wash
- de-ionizing fresh formamide
- prepared new formamide buffers
- prehybe and store at 4C
- Bogdan will take from here
Probes to test on HS cells
- 5 different sized yellow domains
- B02 / B03
- B05 – B07
- B08 – B10
- B10 – B12
- L2D12 (385 kb)
Presentation
- preparing slides for ASBMB meeting
MERFISH
- looking at clustering with new pipeline on old data
Set up new RNAi
- use cells grown for 2 days in complete Schneider’s with FBS
- spin down, resuspend in serum free Schneider’s. (probably should do a PBS wash first next time).
- New RNAi in replicate, on 6 well plates (20 uL each)
- th, rho, water, Pc (v2), ESC, Pc (v2) + ESC + PSC + + SCM
- incubating 50 minutes
Orders
- order new S1 (somehow ran out!?): S1_A647, S2_A647, S3_A750
- first strand synthesis kit arrived today
- Zymo RNA prep kit arrived yesterday (Qiagen RNeasy kit still not arrived).