20.109(S14): TA notes for module 2: Difference between revisions
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'''Key preparation:''' | '''Key preparation:''' | ||
Cells: | Cells: | ||
*Prepare T25 flasks of K1 and xrs6 for every team | *Prepare T25 flasks of K1 and xrs6 for every team | ||
Revision as of 21:57, 11 March 2014
General notes
I need to figure out the quiz dates.
See also GoogleDoc for aliquoting amounts.
Scheme:
- Plate wild type K1 and Ku80 deficient xrs6 cells
- Collect lysates for Westerns to measure Ku80
- Prepare damaged DNA plasmids of varying topologies
- Transfect K1 and xrs6 cells with damaged DNA
- Measure DNA damage repair via NHEJ by flow cytometry
Key preparation:
Cells:
- Prepare T25 flasks of K1 and xrs6 for every team
Westerns:
- Prepare transfer buffer
DNA plasmids:
- Aliquot plasmids
Lipofection: Flow cytometry:
Note to TA:
- While you read the wiki, test each link, and make any link descriptions bold that are not already.
- As you read the protocols, add reagent calculations to the GoogleDoc as needed.
Day-by-day
Day 1
Materials required: None
Day of Lab (T/W):
After Lab:
How it went:
Day 2
Materials required: Please read as much as you can of Parts 1-3 and create an automated worksheet (e.g., in Excel) that will perform the required calculations for the day. A partially prepared worksheet is linked here. We will NOT collect your final worksheet this time; it is only for your benefit.
Day of Lab (R/F):
How it went:
Day 3
Materials required:
Day of Lab (T/W):
After Lab:
How it went:
Day 4
Materials required:
Day of Lab (R/F):
After Lab:
How it went:
Day 5
Materials required:
Day of Lab (T/W):
After Lab:
How it went:
Day 6
Materials required:
Day of Lab (R/F):
After Lab:
How it went:
Day 7
Materials required:
Day of Lab (T/W):
After Lab:
How it went:
Day 8
Materials required:
Day of Lab (R/F):
After Lab:
How it went:
