9:45 A – 8:40P, 9:45P-11:45P
- Submit revised snail kinetics paper.
- Collect virgin Espl/sim cross to TM2/TM6
- Collect virgin Act5 Gal4. Cleared and added new bottle of these to virign collection.
- washout Pc antibody from O/N MTD
- post fix 5 min Pc stained embryos. rinse, move to MeOH, incubate at RT 4 hrs, move to -20C for storage.
DNA FISH
- starting new staining of embryos to test newly synthesized probes
- centromere-dig tagged probe also arrived, see if this works for labeling DNA for embedding.
- diluted cent-dig in 200 uL DNAse free ddH2O. diluted 1:10 in DFISH-hybe buffer, reacting in 100uL.
- Other probes DFISH-cy5 probes v1: Toll, y, sxl, Antp, Ubx, AbdA (listed in order on tube, 100 uL cent-dig is last).
Writing
- Working on BJ review
- Working on figures for BJ review
Confocal
- high background, but dramatically less in nuclei: using nuclear lamina as a boundary, background is quite low and most nuclei have a bright spot, some more than one. Looks promising.
- Still, sometimes hard to tell, especially near nuclear boundary, if a bright spot is real nascent transcription or not.
- substantial varitaion in brightness of signal between probes.
Planning: Fly stocks to order
- BSC 1411: ph[503] — affects both coding units at ph locus. Expression similar to deficiency allele: Df(1)JA52
- 2406 Scm[D1]
- 1728 Pc1/TM1
- Psc — Simon/Bender null alleles show very modest loss of repression. Not available in bloomington. (no small deletions or null mutations available).
- Pcl[D5]
- Pc[XT109] null fails to make protein
- need some good late marked-balancers on chr3: 4432 Ubx-LacZ (should get an anti-Bgal).
- 3516 hs-en (cDNA). is en expression maintained from the native locus?