Tuesday 07/08/14

10:00 am – 8:00 pm, 11:00 pm – 11:50 pm

Sequential staining

  • F06 at least stains this time, some spots quite clear. However staining not very good and STORM pretty weak.
  • Tried flowing more secondary for an extra 2 hours and washing without formamide.
  • stain not much improved.
  • attempting STORM.
  • STORM imaging of previously imaged regions looks horrible.
  • maybe this F06-p1 probe is not good. Test it independently tonight? swap the order of the stain?

Data analysis

  • launched analysis for
    • L3C02,
    • L3E05 and
    • L3D8to12

Troubleshooting continues

  • restain, this time apply S1 O/N. Should answer the question does the F06-P1 probe work. Also added earlier batch of F06-P1
  • also stained each batch of F06-P1 + S1 independently
  • test F06 stain tomorrow morning. If this works I bet we can get BXC to work second (fingers crossed).

Additional new stains

  • D12-P1 (2 uL) + S1 (.5 uL)
  • D11-P1 (2 uL) + S1 (.5 uL)

Project 2

  • see notes
  • screwed some things up in probe making.

Oligo Secondaries

  • playing with scalebars
  • made all images square
  • Need final review of Fig 2 for Brian
  • Need additional images for Supp Fig 14 for Brian.

The other things

Literature

  • reading Helin lab paper on PRC2. Very interesting.
  • proposes PRC2 purely maintenance role, recruited to CpG islands that lack dense nucleosome arrays.

Department mail

  • finally checked this (to pick up rubber cement).
    • one of my IDT shipments went to department mail?! (extra common-primer)
    • Thesis gift from AMOLF
    • nice card from Arnosti group.
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