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]
- Continued from 2014-09-30
- 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.