9:15a – 1:00p, 3:00p-12:00a
Review
- Finalize figure 1.
- Encorporate all figure elements for examples section into figure 2.
- First pass design of new figures complete. Need to apply for permission to reuse images from Raj 2010 Nature, Frankel 2010 Nature,
- start formatting references.
Embryo staining, BX-C, day 2
- 30 min rinse embryos at 60C in 2x SSCT
- rinse embryos at RT in 2x SSCT
- Relabel with 488-WGA
STORM of embryo
- tried imaging in 2x SSC instead of Tris 200mM + 50mM NaCl. Higher background, less good (detectable?) switching.
- Went back to standard Tris buffer. Improved switching but clearly high background and very little 405 response. Really want activator dyes.
Probe making
- Dilute new 20 pmol/uL aliquots of MYcroarray Ubx, AbdA, and AbdB libraries 1:100 in kappa PCR buffer
- PCR mix 1:100 dilution in 1:100 PCR tube and 1:10 PCR tube
- 3 uL of 100 uM primer per 100 uL PCR
- PCR 3+27 rounds, 70C anneal.
- all PCRs look pretty good, no detectable effect of original library dilution