Daniel:Notebook/RNAFISH/2014-12-8
Jump to navigation
Jump to search
RNAFISH
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
- Prepare a cell suspension of 500 000-2 000 000 cells/mL in culture media
- Add 20 uL of cell suspension per dish to the cener; add 80 uL culture media
- Allow cells to adhere for 4 hours at 37 C
- Rinse slides briefly in 1X PBS
- Fix slides for 7 minutes in 4% (v/v) paraformaldehyde in 1X PBS
- Rinse briefly in 1X PBS
- Incubate for 5 minutes in 1 mL of 2X SSCT
- Incubate for 5 minutes in 1 mL of 2X SSCT + 50% (v/v) formamide
- Remove media and add 1 mL of 2X SSCT + 50% (v/v) formamide for storage at 4C; wrap with parafilm
- Slides are typically best used within 1-2 weeks of creation