Daniel:Notebook/HiResChrPaint/2014-2-3
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FISH Slide Prep[edit]
The cancer cells are ready to split, so I made a few more FISH dishes.
Slide Preparation[edit]
1. Prepare a cell suspension of 500 000-2 000 000 cells/mL in RPMI 2. Add 20 uL of cell suspension per dish to the cener; add 80 uL RPMI 3. Allow cells to adhere for 6 hours at 37 C 4. Rinse slides briefly in 1X PBS 5. Fix slides for 5-15 minutes in 4% (v/v) paraformaldehyde in 1X PBS 6. Rinse briefly in 1X PBS 7. Incubate for 5 minutes in 1 mL of 2X SSCT 8. Incubate for 5 minutes in 1 mL of 2X SSCT + 50% (v/v) formamide 9. Remove media and add 1 mL of 2X SSCT + 50% (v/v) formamide for storage at 4C; wrap with parafilm 10. Slides are typically best used within 1-2 weeks of creation