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*[[Hosuk:Lab_Notes|LabNote]] ===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 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=== *Continued from [[Hosuk:LabNotes/2014-9-30|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.
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