Sunday 06/01/14

10:00am – 7:00pm

Goals

  • Call Otto’s order pizza for lab meeting (done)
  • Finish lcPCR and cleanup (done)
  • Set up T7 reactions (done)
  • post-fix Ajaz samples to try immuno-FISH again (done)
  • stain cells (done)
  • finish letter and reply to Nicole about collaboration (done)
  • review and send fellowship progress report to XZ (done)
  • Review manuscript draft for Max (post-poned)

Making probes:

probes

  • B10-B12 (last of the yellows)
  • C09-C12 (last blue, first 2 blacks)
  • D01-D07 (all the end-to-end blacks)
  • [D08-D12 are Bogdan’s internal blacks]

Progress

  • Ran QPCRs
  • Ran PCR cleanup
  • running 30 uL T7 reactions O/N

Cell Staining

  • post-fix in 1% GA 4% PFA 1 hr.
      1. S2 anti-PH
      1. PHwt anti-flag
      1. PHmut anti-flag
  • washed with PBS, then 2x SSC, then prehybe.
  • Stain BXC and ANTC
    • G02-P2-cy3
    • F05-P1-405 (old sample) + S1-488

New Probes to stain

  • E11 + E12 + F01 (G06/alphaTub locus)
  • E11
  • E12
  • F01 (alpha Tub itself)
  • B08
  • B09
  • all with P1-405 + S1-A647

Ph Project

  • remade figure for neg controls (separated channels, matched cropping boxes)
  • sent figures for neg controls to Ajaz
  • review figures
  • finish draft letter for Nicole and sent back
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