Daniel:Notebook/HiResChrPaint/2014-9-19

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FISH[edit]

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Today I'll perform the FISH experiments on the slides I prepared Wednesday

Hybridization[edit]

Sample Matrix

Sample GAD1-488 (pmol) FOXP2-546 (pmol) RBFOX3-594 (pmol) CUX2-647 (pmol) DMD-B-488 (pmol) PPRL-A2-488 (pmol) Cot1-546 (ug)
A1 3.6 (30) X 2.9 (30) X X X X
A2 X 2.9 (30) X 3.3 (30) X X X
B1 X X X X 3.7 (30) X 7.9 (0.4)
B2 X X X X X 21.4 (30) 7.9 (0.4)
  1. Allow stored slides to warm to room temperature
  2. Incubate slides in 2X SSCT +50% (v/v) formamide for 2.5 minutes at 92 C in a pre-warmed coplin jar
  3. Incubate in coplin jar containing 2X SSCT + 50% formamide at 60C for 20 minutes
  4. Remove slides and allow to cool to room temperature
  5. Add 25 uL hybridization cocktail to a 22x22 #1.5 coverslip
  6. Invert slides on to cocktail-covered coverslips; seal with rubber cement
  7. Allow the rubber cement to air-dry for 5 minutes at room temperature
  8. Denature for 2.5 minutes at 92C; keep humid
  9. Transfer slides to a humidified chamber and hybridize overnight at 37C or 42C