Difference between revisions of "20.109(S18): Prep notes for M2"
From Course Wiki
Noreen Lyell (Talk | contribs) (→M2D9) |
Noreen Lyell (Talk | contribs) (→M2D8) |
||
Line 76: | Line 76: | ||
==M2D8== | ==M2D8== | ||
'''24 hr preceding laboratory:''' | '''24 hr preceding laboratory:''' | ||
− | *seed DLD-1 (at | + | *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):''' | '''in tissue culture room (per team):''' |
Revision as of 19:55, 31 January 2018
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
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
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):
- X mL CellTitre Glo reagent
- Y mL PBS
- Z mL media (without FBS, antibiotics)