Daniel:Notebook/RNAFISH/2014-12-8

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Revision as of 23:55, 8 December 2014 by >Djacobse (Created page with "=RNAFISH= Back to Calendar Based off the cell line analysis from last week, I've concluded that investigati...")
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RNAFISH

Back to Calendar

Based off the cell line analysis from last week, I've concluded that investigating CUX2 and GAD1 in the BE2-C and U87-MG cell lines (respectively) should yield some pretty good results. First, I need to prep my samples.

Dish Preparation

  1. Prepare a cell suspension of 500 000-2 000 000 cells/mL in culture media
  2. Add 20 uL of cell suspension per dish to the cener; add 80 uL culture media
  3. Allow cells to adhere for 4 hours at 37 C
  4. Rinse slides briefly in 1X PBS
  5. Fix slides for 7 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