cell staining
- control worked, 50kbp probe did not. However, 50kbp probe does show weak global nuclear staining immediately after removing probe, which is not visible in control channels.
- hot washes started
- 60C SSCT hot washes dramatically reduce signal from control channel. Try 37C?
Probe making
- forgot to heat denature, running as a single band 🙁 Will need to extract and repeat.
Fly work
- never made it to morning virgin collections.
- skipped evening collections
embryo in situs
- day 3, washing out primary
- secondary staining
PCR screening
- L3R9, L7R12,L5R4,L8R10, Espl, then Sim
- 3A LongAmp, 59C anneal, 10min elong, 32 cycles
- 3B LongAmp, 54C anneal, 10 min elong, 32 cycles
- 4A BioXAct, 59C anneal, 5:30 69C elong, 32 cycles
- 4B BioXAct, 63C anneal, 5:00 69C elong, 32 cycles