Sunday 09/01/13

8:00a – 11:00p

Review

  • working on incorporating feedback from Jeff into revision
  • added new figure on explanation of Markov Models

Cell staining

  • goals: test probes from new library. Test Tye665 dye
  • samples: Bxc-405 + Tye665 on clone8. Bxc-405 + cy5 on clone8.
  • G2, G3, G4, G5, G6 and G8 on Kc167 cells. + no primary control.
  • dilutions: 24 uM Bxc-405. 25 picomol/uL -> 1 uL. Tye665 and cy5 secondaries (at 100 uM) dilute 1:5 in hybe buffer and add 1 uL per reaction.
  • Raw G2-G8 concentrations are 400-1000 ng/uL. Add 5uL probe + 1 uL cy5-secondary-dock + 1 uL 405 diluted tertiary.
  • denature at 78C (heat block set 85C). Hybe O/N at 47C.
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