Saturday 02/08/14

10:30a – 4:45p, 10:00p-11:50p

Chromatin

Deep sequencing of libraries

  • finish gel extraction aborted last night due to mini-centrifuge failure.

Data analysis

  • Analyze new ANTP data
  • issue: need to fix integrated drift correction, have some weird errors
  • interpolation was not correct — need to convert bead frames to image frames.
  • drift correction now working smoothly on integrated bead data
  • issue 2: contrast averaging is too a bit too strong.
    • Contrast is good but images are getting compressed into very low bit-depth relative to available dynamic range.
    • maybe we should integrate over 60 frames and divide by 20 instead of 60? Lots of bead frames can in fact be added together without risking saturation.

Ph Project

Confocal Imaging of S2 cells

Prep

  • make fresh Glox buffer solution
  • let’s see if we can get this to work in chambered slides with Glox buffer.

Samples imaged

  • Ph-Flag-WT
  • Ph-Flag-M
  • Endog. Ph in WT
  • Endog. Ph in M
  • Psc in WT
  • Pc in WT
  • Psc in M
  • Pc in M

Observations

  • Ph-Flag staining generally much weaker than others. Not sure why, seemed fine by STORM.
  • With the usual contrast issues of confocal, there is some evidence for bodies in all samples.
    • Most pronounced in Ph-FLAG-WT.
    • Pc-WT vs mutant also potentional difference.

Fly work

  • flipped stocks.
  • All survived, some have dry food debris carry over but hopefully will recover.
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