Difference between revisions of "20.109(F19): Prep notes for M1"
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Noreen Lyell (Talk | contribs) (→M1D4) |
Noreen Lyell (Talk | contribs) (→M1D1) |
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'''48 hr preceding laboratory:''' | '''48 hr preceding laboratory:''' | ||
*seed CHO (350K) in T75 flasks, 1 per team | *seed CHO (350K) in T75 flasks, 1 per team | ||
− | *[[cell expansion schedule]] for D1, D3, and D4 prep | + | *[[Media:Fa19 M1 cell seeding schedule.xlsx| cell expansion schedule]] for D1, D3, and D4 prep |
'''in tissue culture room (per team):''' | '''in tissue culture room (per team):''' |
Latest revision as of 17:48, 4 September 2019
M1D1
48 hr preceding laboratory:
- seed CHO (350K) in T75 flasks, 1 per team
- cell expansion schedule for D1, D3, and D4 prep
in tissue culture room (per team):
- 1 fresh 12-well plate
- 5.5 mL PBS
- 2.5 mL trypsin
- 35 mL media
- not aliquoted: trypan blue
in main lab (per team):
- none
M1D2
in tissue culture room (per team):
- 135 μL 12 mM MMS
- 8 μL 100 mM As
- 45 mL PBS
- 40 mL media
- not aliquoted: EdU
in main lab (per team):
- not aliquoted: NMP stock, 1X PBS, gel bond film (pre-cut), markers, rulers
M1D3
48 hr preceding laboratory:
- seed CHO (350K) in T75 flasks, 1 per team
- teaching faculty will harvest cells just prior to student use and concentrate to 500K / mL
prior to laboratory
- teaching faculty will prepare Click-iT reaction mix
in tissue culture room (per team):
- none
in main lab (per team):
- 9 mL 1X TBS
- not aliquoted: Triton-X, BSA, anti-γH2AX antibody, mounting media, 1X PBS, LMP solution
students will prepare following solutions in class:
- Triton-X solution = 2.5 mL 1X TBS containing 0.2% Triton-X
- blocking solution = 2.5 mL 1X TBS containing 1% BSA
- 1° antibody solution = 2.5 mL blocking solution containing 1:1000 dilution of anti-γH2AX antibody
M1D4
48 hr preceding laboratory:
- seed CHO (350K) in T75 flasks, 1 per team
- teaching faculty will harvest cells just prior to student use and concentrate to 500K / mL
prior to laboratory
- teaching faculty will prepare CometChips, 1 per team
in tissue culture room (per team):
- none
in main lab (per team):
- 50 μL 12 mM MMS
- 10 μL 100 mM As
- 12 mL media
- not aliquoted: 1X PBS, LMP solution, alkaline lysis buffer
M1D5
in tissue culture (per team):
- none
in main lab (per team):
- not aliquoted: unwinding / electrophoresis buffer, neutralization buffer, SYBR gold
M1D6
in tissue culture room (per team):
- none
in main lab (per team):
- none
M1D7
in tissue culture room (per team):
- none
in main lab (per team):
- none