Daniel:Notebook/RNAFACS/2015-10-1
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Nuceli Adhesion[edit]
Since the mouse trial (yesterday) went well I am going to try out some human nuclei.
- Clean glass slide in 100% EtOH; allow to air dry
- Remove cells from -80C and spin down for 5 min at 1000g
- Remove supernatant, careful not to disturb the pellet
- Add 500uL of 4% paraformaldehyde to the tube and wait 5 min
- Centrifuge cells for 5 min at 1000g
- Remove supernatant, careful not to disturb the pellet
- Add 500 uL of 70% EtOH
- Centrifuge cells for 5 min at 1000g
- Remove supernatant, careful not to disturb the pellet
- Resuspend nuclei in 20 uL 70% EtOH
- Deposit on to glass slide
- Use a coverslip to gently spread the 20 uL across the slide
- Be careful not to scratch the slide
- Dry the slide using a heating block at 60C for 10-15 min
- Add 15 uL Slowfade + DAPI and seal with a coverslip