20.109(S14): TA notes for module 2

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

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After Lab:

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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):

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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

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Day 8

Materials required:

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