20.109(S14): TA notes for module 2

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(General notes)
(General notes)
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'''Key preparation:'''
'''Key preparation:'''
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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 00:57, 12 March 2014

20.109(S14): Laboratory Fundamentals of Biological Engineering

Home        Schedule Spring 2014        Assignments       
Module 1        Module 2        Module 3              

Contents

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:
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