Difference between revisions of "20.109(F18): Prep notes for M1"
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Noreen Lyell (Talk | contribs) (→M1D6) |
Noreen Lyell (Talk | contribs) (→M1D5) |
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'''in tissue culture (per team):''' | '''in tissue culture (per team):''' | ||
*50 mL media | *50 mL media | ||
+ | *40 mL PBS | ||
*not aliquoted: H<sub>2</sub>O<sub>2</sub> (on ice at center bench) | *not aliquoted: H<sub>2</sub>O<sub>2</sub> (on ice at center bench) | ||
Line 68: | Line 69: | ||
*not aliquoted: Triton-X, BSA, anti-γH2AX antibody | *not aliquoted: Triton-X, BSA, anti-γH2AX antibody | ||
− | ''students will prepare following solutions in class'' | + | ''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 | |
− | + | ||
− | + | ||
− | + | ||
==M1D7== | ==M1D7== |
Latest revision as of 18:50, 26 September 2018
M1D1
48 hr preceding laboratory:
- seed M059K (at 500K) and M059J (at 500K) in T25 flasks, 1 per team
- cell expansion schedule for D1, D3, and D4 prep
in tissue culture room (per team):
- 2 fresh T75 flasks
- 4.5 mL PBS
- 1.5 mL trypsin
- 25 mL media
in main lab (per team):
- not aliquoted: NMP stock, 1X PBS, gel bond film (pre-cut), markers, rulers
M1D2
- students will use cells seeded M1D1 for CometChip loading experiment
in tissue culture room (per team):
- 11 mL PBS
- 5 mL trypsin
- 11 mL media
- not aliquoted: trypan blue
in main lab (per team):
- not aliquoted: LMP solution
M1D3
48 hr preceding laboratory:
- seed M059K (at 1M) and M059J (at 1M) in T75 flasks, 1 per team
- teaching faculty will harvest cells just prior to student use and concentrate to 500K / mL
in tissue culture room (per team):
- none
in main lab (per team):
- not aliquoted: 1X PBS, LMP solution, H2O2 (on ice at front bench), alkaline lysis buffer
M1D4
48 hr preceding laboratory:
- seed M059K (at 500K) and M059J (at 500K) in T75 flasks, 1 per team
in tissue culture room (per team):
- 11 mL PBS
- 5 mL trypsin
- 50 mL media
in main lab (per team):
- not aliquoted: unwinding / electrophoresis buffer, neutralization buffer
M1D5
in tissue culture (per team):
- 50 mL media
- 40 mL PBS
- not aliquoted: H2O2 (on ice at center bench)
in main lab (per team):
- none
M1D6
in tissue culture room (per team):
- none
in main lab (per team):
- 9 mL 1X TBS
- not aliquoted: Triton-X, BSA, anti-γH2AX antibody
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
M1D7
in tissue culture room (per team):
- none
in main lab (per team):
- none