Daniel:Notebook/HiResChrPaint/2014-2-3

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FISH Slide Prep[edit]

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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