Friday 08/14/15

9:00 am – 5:45 pm


  • need to do fewer review experiments at once so we can do them more carefully
  • should focus on the live imaging and Pc analysis
  • need to get good QPCR
  • need to set up deep-sequencing
  • need to analyze internal domains
  • need to do 2-color experiments (try L2 G05 G06).

Lab meeting

  • Jiang practice talk (see notes)


  • started from RNA again, ran total RNA prep using oligoT primer
  • spiked in 2 uL RNase H as per the SS3 kit, incubated at 37
  • set up QPCR reactions
    • diluted DNA 1:5, then added 2.5 uL per tube.
  • revised quality controls on QPCR analysis script
  • most genes no amplification
    • Dfd amplified inall the Ph KD but none of the controls
  • probably not enough cDNA (or maybe inhibitor?)

Meeting with XZ

  • live imaging plans:
    • move forward with analysis of SIM data
    • focus on the TIRF2-telo
  • ph
    • internal domain analysis
    • domain cross-boundary interaction
    • qPCR
    • sequencing
    • immuno-staining (?)
    • western (?)
  • other things are lower priority.

TIRF2 imaging

  • test imaging TIRF2-GFP fusion
  • these look very encouraging
  • clearly visible on Turnkey by eye
  • lots of spots, low nuclear background, clear signal.
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