20.109(S14): TA notes for module 2: Difference between revisions
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===Day 1=== | ===Day 1=== | ||
'''Materials required:''' | '''Materials required:''' | ||
1. Aliquots of PBS, CHO media, and 0.25% Phenol Red Trypsin for each group | |||
2. T25 flask of K1 and xrs6 cells for each group | |||
*Flasks for next day seeded at 600,000 cells | |||
*Flasks for day after next, seeded at 400,000 cells | |||
*Flasks for 3 days in future, seeded at 200,000 cells | |||
'''Day of Lab (T/W):''' | '''Day of Lab (T/W):''' | ||
*No Quiz | |||
*Prepare remaining aliquots for students | |||
*Warm up media, PBS, and trypsin half hour before students use reagents | |||
Additional Prep.: | |||
*Digest pMAX-BFP-2MCS2 for testing damage topologies | |||
*Pour gel for purification | |||
'''After Lab:''' | '''After Lab:''' | ||
*Aliquot new shipment of trypsin | |||
'''How it went:''' | |||
Day went pretty smoothly; not many questions on Part 4 exercise. | |||
===Day 2=== | ===Day 2=== | ||
Revision as of 22:06, 11 March 2014
General notes
I need to figure out the quiz dates.
See also GoogleDoc for aliquoting amounts.
Scheme: Each pair of students will test K1 and xrs6 cells for Ku80 production. They will then assess the ability of K1, xrs6, and DNA-PKcs inhibited cells to repair DNA damage via plasmid based assay and 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:
1. Aliquots of PBS, CHO media, and 0.25% Phenol Red Trypsin for each group 2. T25 flask of K1 and xrs6 cells for each group
- Flasks for next day seeded at 600,000 cells
- Flasks for day after next, seeded at 400,000 cells
- Flasks for 3 days in future, seeded at 200,000 cells
Day of Lab (T/W):
- No Quiz
- Prepare remaining aliquots for students
- Warm up media, PBS, and trypsin half hour before students use reagents
Additional Prep.:
- Digest pMAX-BFP-2MCS2 for testing damage topologies
- Pour gel for purification
After Lab:
- Aliquot new shipment of trypsin
How it went:
Day went pretty smoothly; not many questions on Part 4 exercise.
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
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Day 4
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Day 5
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Day 6
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Day 7
Materials required:
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Day 8
Materials required:
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