Wednesday 10/31/12

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.
  • 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).  
  • Working on BJ review
  • Working on figures for BJ review
  •  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?
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