Daniel:Notebook/HiResChrPaint/2014-6-29

From ZhangLabWiki
Revision as of 22:16, 30 June 2014 by >Djacobse (Created page with "=FISH 70% Formamdide= Back to Calendar ==Slide Preparation== #Prepare a cell suspension of 500 000-2 000 000 cells/mL in RPMI #Add 20 uL o...")
(diff) ← Older revision | Latest revision (diff) | Newer revision → (diff)
Jump to navigation Jump to search

FISH 70% Formamdide

Back to Calendar

Slide Preparation

  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