Altman:Protocols/TC Protocols/Freezing SF9

From OpenWetWare

Jump to: navigation, search


Department of Physics, Willamette University

'


Home        Contact        Lab Members        Publications        Research        Protocols        Pictures       


Freezing down SF9 cells for long-term storage

1. Grow the cells to a density that is greater than or equal to 1*10^7 viable cells/mL

2. Pellet the cells by centrifugation: place the 35 mL of cells in a 50-mL conical, and spin at 1000 rpm for 10 minutes

3. Remove the media; this media is "conditioned media"

4. Combine the conditioned media 1:1 with fresh media

5. Add DMSO to the media to a final concentration of 7.5% (e.g. 2.6 mL DMSO + 35 mL media)

6. Resuspend the cells in the media+DMSO to return the cells to a final density of greater than or equal to 1*10^7 viable cells/mL

7. Aliquot 1.5 mL of cells into 2.0 mL cryovials

8. Place tubes between two pieces of styrofoam and place in a -20 freezer overnight

9. Place in liquid nitrogen for long term storage
Personal tools