Daniel:Notebook/HiResChrPaint/2014-6-29: Difference between revisions
Jump to navigation
Jump to search
>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...") |
>Djacobse No edit summary |
||
(2 intermediate revisions by the same user not shown) | |||
Line 2: | Line 2: | ||
[[Daniel:Notebook/HiResChrPaint|Back to Calendar]] | [[Daniel:Notebook/HiResChrPaint|Back to Calendar]] | ||
Prepared 5 dishes using following protocol. | |||
*Sample A** has 80/20 cells/RPMI | |||
*Samples A,B,C,D have 20/80 cells/RPMI | |||
==Slide Preparation== | ==Slide Preparation== | ||
Line 15: | Line 19: | ||
#Remove media and add 1 mL of 2X SSCT + 50% (v/v) formamide for storage at 4C; wrap with parafilm | #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 | #Slides are typically best used within 1-2 weeks of creation | ||
[[Category:HRCP]] [[Category:Slide Preperation]] [[Category:20140630]] |
Latest revision as of 17:37, 2 July 2014
FISH 70% Formamdide[edit]
Prepared 5 dishes using following protocol.
- Sample A** has 80/20 cells/RPMI
- Samples A,B,C,D have 20/80 cells/RPMI
Slide Preparation[edit]
- Prepare a cell suspension of 500 000-2 000 000 cells/mL in RPMI
- Add 20 uL of cell suspension per dish to the cener; add 80 uL RPMI
- Allow cells to adhere for 6 hours at 37 C
- Rinse slides briefly in 1X PBS
- Fix slides for 5-15 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