10:00A – 11:30P
- coating 5o mm coverslips
- put coverslips in metal, vertical mount holder. Don’t stay balanced in ceramic holder. Need to alternate positions or the coverslips stick.
- Do coating in 400 mL in 1000 mL beaker (not 300 mL in 600 mL beaker, metal rack does not fit).
- Sonication steps need to be done in square glass case (only thing that fits holder and fits in sonicator).
- Ultra-sectioning HP-6, H3-750 sample.
- stuff growing in SOC. explains colonies in control ligation possibly. Try transforming again…
- repeat transformations using all new pipettes and new sterile tubes, new SOC.
- gel extracting PCRs 2,3,5, 8 (no well/gel-box combo capable of 50 wells). Largest is 8 20 uL wells, so we extract 4.
- concentrations of Dfd2, AbdB-1, AbdB-3, and iab5E all look good: 12-20 ng/uL.
- Ligating O/N at 4C with 2 uL plasmid: 1 uL PGEM.
- cut sections: 3 slides of H3-750, Hp1-647.
- 4 slides of H3-647, Hp1-750. ribbons not holding together well. humidity low and sections curling in and swirling with extra static electricity.
- working on journal club presentation on Li and Pirrotta 2011 paper.
- Fixed Pc-GFP flies 15 min fix, prepped for oligo-STORM
- hybridizing with oligo-paint Cy5 probe. embryos translucent in hybe, very hard to see in small volume PCR tube. Denatured probe and embryos separately, 5 min, then together 3 min (at 94C), then dropped to 37C. Embryos at RT while heat block drops temp.
- Oligo paint probe 50 picomoles in 50 uL.
- Select small number of embryos to mount after quick hybe rinse at RT, 2x SSC dilution, then mount.