AlanFung:LabNotes/Rolony/2013-9-26: Difference between revisions

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>Alan6017518
>Alan6017518
Line 9: Line 9:
*Add in 2X ssc
*Add in 2X ssc
*image
*image
==Matlab Image Analysis==

Revision as of 21:35, 26 September 2013

Strip off dye coupling with formamide

  • Aspirate PBS from mattek dish with primary/secondary rolonies
  • Add 200ul preheated (65C-80C) 80% formamide in 2X SSC
  • Incubate 10min RT
  • Rinse w H2O
  • Add in 1:200-1uM 200ul (preheat 65C) ACTB-primary rolony target probe (Alexa 488) (200uM ACTB_1stRolonyFISH)
  • Incubate 10min RT
  • Rinse w 2x SSC twice
  • Add in 2X ssc
  • image

Matlab Image Analysis