Thursday 12-01-11

10:30 A — 11:00P

  • Testing WGA labeling after Xylene treatment with WGA in PBS and WGA in block.
  • Steve Chu lecture
  • Analyzing new MP08, MP05 data.
  • Running new MP06 data
  • apparently WGA in block does not bind substantially to membranes.  Should use without block, after secondaries.  More washing would help lower background.
  • confocal imaging of WGA-488, en-555, H3K27-647 sectioned embryos: resin still there, en looks beautiful, WGA detectable, H3K27 very weak, probably due to in part to resin?
  • sodium ethoxide disolving resin does not always work.  Seems like more thoroughly washed sections have mechanically removed resin.  When you can see by eye the square blocks still there after washing, they probably are still there.  Longer treatment in new sodium ethoxide.
  • Control testing STORM imaged H3K4 — embryo sections imaged don’t have confocal detectable H3K27 647 staining.  Some other sections do, try these.
  • reading about engrailed regulation — good long range enhancers.  Functional PREs in both pairing and silencing tests.  Why do some en cells stay on and others stay off?  en promoters preferentially land near the en gene (within 300 kb).
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