Daniel:Notebook/HiResChrPaint/2014-1-22

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Revision as of 00:35, 23 January 2014 by >Djacobse (Created page with "=FISH= Back to Calendar Prepping some samples to hopefully use with Alu positive control by the weekend. ==FISH Dish Preparation== 1. Pr...")
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FISH

Back to Calendar

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