Daniel:Notebook/HiResChrPaint/2014-1-22
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FISH
Prepping some samples to hopefully use with Alu positive control by the weekend.
FISH Dish Preparation
1. Prepare a cell suspension of 500 000-2 000 000 cells/mL in growth media 2. Add 100 uL of cell suspension per slide roughly in the center 3. Allow cells to adhere for 1-3 hours at the growth temp of the cell line 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 2X SSCT 8. Incubate for 5 minutes in 2X SSCT + 50% (v/v) formamide 9. Replace with fresh 2X SSCT + 50% (v/v) formamide for storage at 4C 10. Slides are typically best used within 1-2 weeks of creation
- For cancer cell line (NCIH 1975) I'd recommend ~10 uL cell suspension and 90 uL DMEM