9:00 am
pairing meeting
- rehearse talk (8:45-9:15)
- see notes
Experiments: QPCR for RNAi (9:15 am – 11:05 am)
- RNA concentrations from yesterday: 600 – 1200 ng/uL (55 uL volume)
- ran RNase H digestion last night, at 4C in block O/N.
- ran clean up column (DCC5)
- cDNA concentrations from polyT primed RT reaction: 40 – 100 ng/uL
Setting up QPCR
- 40x master mix (9 primer pairs, 4 samples)
- per each (total for master)
- 12.5 uL Hot-start Phusion mix (500 uL)
- 2.5 uL Eva Green (50 uL)
- 6 uL ddH2O (240 uL)
- 2 uL cDNA
- 2 uL primer
- plate layout
- col 1:11 = Act, Tbu, Gapdh, ubx, Abd-B, Dfd, Antp, en, ph-p, ph-d1, ph-d2,
- rows 1:4 = PhWT 1 10-16, PhWT 2 10-16, Wt 2 day 10-22, Ph 2 day 10-22.