Difference between revisions of "Freezing and Thawing S2 Cells"
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| + | ==SOP== | ||
| + | *[[SOP- Cryogenic Materials]] | ||
| + | *[[SOP- Biosafety Cabinets]] | ||
| + | |||
| + | ==METHOD== | ||
#Use and early passage (1-10) of exponentially growing (about ~10^7 cells). | #Use and early passage (1-10) of exponentially growing (about ~10^7 cells). | ||
#Detach by pipeting a stream of media over the cells | #Detach by pipeting a stream of media over the cells | ||
Latest revision as of 14:28, 5 June 2020
SOP
METHOD
- Use and early passage (1-10) of exponentially growing (about ~10^7 cells).
- Detach by pipeting a stream of media over the cells
- Transfer into a 15 mL falcon tube
- Spin at 500g for 10 min, save the supernatant
- Prepare freezing media:
- Add 4.5 mL supernatant
- Add 4.5 mL fresh media
- Add 100 uL sterile DMSO
- Resuspend to 1/10th original culture volume
- Dispense 0.5 mL into sterile cryovials
- Place in isopropanol freezing chamber and store at -80 for about a day
- Transfer to liquid nitrogen
- Cells should be viable for >5 years
see PMID 18388942
