Difference between revisions of "20.109(F19): Prep notes for M3"
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==M3D1== | ==M3D1== | ||
+ | |||
+ | '''24 hr preceding laboratory:''' | ||
+ | *seed MEF (250K) in T25 flasks, 1 per team | ||
+ | *[[Media:Fa19 M3 cell seeding schedule.xlsx | 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, dH<sub>2</sub>O | ||
+ | |||
+ | ==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 |
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