20.109(S18): Prep notes for M2
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Revision as of 19:14, 16 February 2018 by Noreen Lyell (Talk | contribs)
M2D1
48 hr preceding laboratory:
- seed DLD-1 (at 250K) and BRCA2-/2 (at 500K) in T25 flasks, 1 per team
- cell expansion schedule for D1, D2, and D8 prep
in tissue culture room (per team):
- 2 fresh T25 flasks
- 5 mL PBS
- 3 mL trypsin
- 12 mL media
in main lab (per team):
- none
M2D2
24 hr preceding laboratory:
- seed DLD-1 (at 500K) and BRCA2-/2 (at 1M) in T75 flasks, 2 per team
in tissue culture room (per team):
- 22 mL PBS
- 20 mL media (for etoposide treatment)
- 25 μL etoposide
- 20 mL media (for drug treatment)
- 10 μL loperamide
- 10 μL mibefradil
in main lab (per team):
- none
M2D3
prior to laboratory:
- prepare aliquots of purified RNA to supplement student samples
in tissue culture room (per team):
- 40 mL PBS
- 10 mL trypsin
in main lab (per team):
- RNA purification aliquots
- cDNA aliquots
M2D4
in tissue culture room (per team):
- none
in main lab (per team):
- none
M2D5
prior to laboratory:
- prepare aliquots of cDNA to supplement student samples
in tissue culture room (per team):
- none
in main lab (per team):
- X mL RNase free water
- qPCR aliquots
M2D6
in tissue culture room (per team):
- none
in main lab (per team):
- none
M2D7
in tissue culture room (per team):
- none
in main lab (per team):
- none
M2D8
24 hr preceding laboratory:
- seed DLD-1 (6 wells at 25K) and BRCA2-/2 (6 wells at 50K) in 12-well plate, 1 per team
in tissue culture room (per team):
- 28 mL PBS
- 18 mL media (for etoposide treatment)
- 20 μL etoposide
- 50 mL media (for drug treatment)
- 10 μL loperamide
- 10 μL mibefradil
in main lab (per team):
- none
M2D9
in tissue culture room (per team):
- none
in main lab (per team):
- 6.5 mL CellTitre Glo reagent
- 30 mL PBS
- 6.5 mL media