Difference between revisions of "20.109(F19): Prep notes for M3"
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Noreen Lyell (Talk | contribs) (→M3D2) |
Noreen Lyell (Talk | contribs) (→M3D2) |
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'''in tissue culture room (per team):''' | '''in tissue culture room (per team):''' | ||
*25 mL PBS | *25 mL PBS | ||
− | *25 mL media | + | *25 mL serum-free media |
*not aliquoted: ligands | *not aliquoted: ligands | ||
'''in main lab (per team):''' | '''in main lab (per team):''' | ||
− | * | + | *15 mL PBS |
− | + | ||
*5 mL trypsin | *5 mL trypsin | ||
− | *15 | + | *15 media |
− | *not aliquoted: protease inhibitor | + | *1.5 mL PBS containing protease inhibitor |
+ | *not aliquoted: protease inhibitor | ||
==M3D3== | ==M3D3== |
Latest revision as of 18:44, 1 November 2019
M3D1
24 hr preceding laboratory:
- seed MEF (250K) in T25 flasks, 1 per team
- cell expansion schedule for D1 and D2 prep
in tissue culture room (per team):
- 1 fresh T75 flask
- 3.5 mL PBS
- 1.5 mL trypsin
- 15 mL media
- not aliquoted: trypan blue
in main lab (per team):
- none
M3D2
48 hr preceding laboratory:
- seed MEF (2-3M) in T75 flasks, 4 per team
in tissue culture room (per team):
- 25 mL PBS
- 25 mL serum-free media
- not aliquoted: ligands
in main lab (per team):
- 15 mL PBS
- 5 mL trypsin
- 15 media
- 1.5 mL PBS containing protease inhibitor
- not aliquoted: protease inhibitor
M3D3
in tissue culture room (per team):
- none
in main lab (per team):
- 75 μL Laemmli sample buffer
- 30 mL blocking buffer
- not aliquoted: molecular weight standards, TGS running buffer, dH2O
M3D4
in tissue culture room (per team):
- none
in main lab (per team):
- 10 mL blocking buffer (2 per team)
- not aliquoted: TBS-T
M3D5
in tissue culture room (per team):
- none
in main lab (per team):
- none