9:05 am – 7:30 pm, 8:15 pm – 11:40 pm
- testing control L4E24 embryos. stains look great.
- compare to L7 embryos — no staining.
- ran series removing S2 with toe-hold 5 bp. Looks excellent.
- Gel tests
- try to anneal probe and secondary on bench, no cells. Test if still bound on gel.
- 5% PAGE gel. ran too long and gel was a bit too old
- smears suggest partially degraded RNA slowing down template to various degrees
- found the likely problem — 8 mM NaOH instead of 1 M NaOH in the RNA degradation step.
- re-ran digestion, re-ran probe clean-up
- re-ran hybridization
- S1rc + L7 common = GCGATGGTAGACGGCGTATGAATTCGGCAGAC GACCCGTCAGGATAGGCCCT
Microscope building prep
- requesting quotes from MPB, Nikon, Coherent, and Newport
- write microscope justification
- book flights back from San Fran
- diagnostic gel
- detailed labeling of samples