Hosuk:LabNotes/2014-8-19

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Revision as of 18:46, 22 August 2014 by >Hosuki78 (Created page with "*LabNote ===Decoding with Agi26k0gap Padlock Probe=== *Human brain tissue from Yun **HB3, section 9 **Total 15 tissue sections *Mouse tissue : Total 15 t...")
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Decoding with Agi26k0gap Padlock Probe

  • Human brain tissue from Yun
    • HB3, section 9
    • Total 15 tissue sections
  • Mouse tissue : Total 15 tissue sections
  • Stored in -80C


Procedure

Fixation
  • 4% Formaldehyde : 2.5mL 16% Paraformaldehyde + 1mL 10x DEPC-PBS + 6.5mL DEPC-H2O (Tot. 10mL)
    • 16% Paraformaldehyde Solution)
    • Manufacture : Electron Microscopy Diatome --> 16% PARAFORMALDEHYDE AQ SOLUTN 50-980-486, No.:15700
    • Supply : (Fisher Scientific), 50980486
      • Paraformaldehyde - methanol free solution. A more efficient and rapid penetrant fixative used in combination with Glutaraldehyde and Acrolein fixatives. Easy to break, prescored, 10 ml ampoules sealed under inert gas 10 ml
  • Take Tissue glass slide from -80C, and put them to room temp. for 2min.
  • Add 4% formamide on a tissue, incubate for 30min at room temp.
  • Wash with PBS 3 times (add PBS in 3 wells of 6 well plate and series washing with glasses)
  • Attach glass with glue
    • Cyanoacrylate adhesives : High-viscosity, Sigma-Aldrich, Z105902-1EA
    • Wipe PBS from the glass (around tissue)
    • Apply glue along the edge of the glass, put on the bottom of MatTek dish, wait for 5min

File:TissueGlassOnMatTek 2.png File:TissueGlassOnMatTek 1.png



Preparation
Rolony Process
Reverse Transcription

Summary of samples