Hosuk:LabNotes/2014-10-1: Difference between revisions

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===Tissue Section : fixing test===
*Continued from [[Hosuk:LabNotes/2014-9-30|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 tissue, )
**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
*RCA at 2014-10-02
===ppCOL1A1 + Suppressor test===
*Continued from [[Hosuk:LabNotes/2014-9-30|2014-09-30]]
*Continued from [[Hosuk:LabNotes/2014-9-30|2014-09-30]]



Revision as of 17:33, 2 October 2014


Tissue Section : fixing test

  • 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 tissue, )
    • 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
  • RCA at 2014-10-02


ppCOL1A1 + Suppressor test


  • 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.