20.109(S14): TA notes for module 2

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(General notes)
(Day 1)
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===Day 1===
===Day 1===
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'''Materials required:''' None
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'''Materials required:'''  
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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:'''
-
 
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*Aliquot new shipment of trypsin
'''How it went:'''
'''How it went:'''
 +
 +
Day went pretty smoothly; not many questions on Part 4 exercise.
===Day 2===
===Day 2===

Revision as of 01:06, 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: 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

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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