9:50 pm – 11:45 pm
RNAi
RNA prep
- set up T7 reactions
- 10 x 9 = 90 uL NTP buffer mix
- 2 x 9 = 18 uL T7 pol mix
- 2 x 9 = 18 uL RNasin
- 6 x 9 = 54 uL ddH2O
- add to 10 uL of template DNA per well.
- incubating at 37 C for 10 hrs O/N
RNAi treatment
- add RNAi in 12 well plates
- 10 uL of ~800 ug/uL RNA samples from BB.
- 750 uL of cells resupended in serum free Schneider’s media (from SFX)
- setup (top left to bottom right)
- Th (two wells)
- rho (two wells)
- water (two wells)
- RNAs 1-3 (one well)
- RNAs 5-6 (one well)
- Pc (one well)
- Psc (one well)
Cell culture
- passaged cells to a new plate of SFX
- moved some cells to a new plate of Schneider’s media + 10% FBS