Daniel:Notebook/HiResChrPaint/2014-9-19
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FISH[edit]
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) |
- Allow stored slides to warm to room temperature
- Incubate slides in 2X SSCT +50% (v/v) formamide for 2.5 minutes at 92 C in a pre-warmed coplin jar
- Incubate in coplin jar containing 2X SSCT + 50% formamide at 60C for 20 minutes
- Remove slides and allow to cool to room temperature
- Add 25 uL hybridization cocktail to a 22x22 #1.5 coverslip
- Invert slides on to cocktail-covered coverslips; seal with rubber cement
- Allow the rubber cement to air-dry for 5 minutes at room temperature
- Denature for 2.5 minutes at 92C; keep humid
- Transfer slides to a humidified chamber and hybridize overnight at 37C or 42C