Hosuk:LabNotes/2014-10-1

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Tissue Section : fixing test[edit]

  • Continued from 2014-09-30
  • Result : Tissue remained well, but not many rolonies…Need more permeablization?


  • Prepare another run
    • S1 : Mouse brain tissue, 4hr fix, Pepsin for 20min
    • S2 : Mouse brain tissue, 4hr fix, Pepsin for 10min (--> fail to prepare sample with this tissue, so it will not be used for this time)
    • S3 : Mouse brain tissue, 2hr fix, Pepsin for 20min
    • S4 : Mouse brain tissue, 2hr fix, Pepsin for 10min
    • And PGP1F sample
  • --> Therefore I have 4 MatTek dishes (S1, S3, S4 : Mouse Brain Tissue, and 1 PGP1F)


  • Start fix and RT at 2014-10-01


ppCOL1A1 + Suppressor test[edit]


  • Wash with 1X PBS twice
  • Add 200ul 0.5uM FISSEQ_ppRCA in 2X SSC and incubate at 60C for 30min
  • Wash with 2X SSC twice
  • Add 200ul RCA Mix and incubate at 30C for 16hrs
    • H2O : 172uL
    • 10x Buffer : 20uL
    • dNTP : 2uL
    • a-dUTP : 4uL
    • Phi29 (1000 U/µl) : 0.5uL
  • Incubate at 30C for 20hr.