20.109(S14):Data analysis (Day7): Difference between revisions
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Topic 4: but the most interesting/fun will be flow analysis: mean vs median choice; breaking down the Day 5 equation a bit more | Topic 4: but the most interesting/fun will be flow analysis: mean vs median choice; breaking down the Day 5 equation a bit more | ||
Oh, and just a word about the MCS/GC/etc. issue for context | |||
==Protocols== | ==Protocols== |
Revision as of 09:47, 4 April 2014
Introduction
Topic 1: document some of the lecture info on NHEJ key players, leading to
Topic 2: more about C401 inhibitor, and
Topic 3: more about colony-forming assay and staining approach
Topic 4: but the most interesting/fun will be flow analysis: mean vs median choice; breaking down the Day 5 equation a bit more
Oh, and just a word about the MCS/GC/etc. issue for context
Protocols
Part 1: Stain irradiated cell colonies
Option to do it on M3D1 if they want to grow longer for bigger colonies?
All in main lab: rinse w/ 2mL pre-warmed PBS, add 2 mL Coomassie for 1 hr w/shaking, save it afterward, rinse w/PBS again, let dry a little bit, then count colonies right away. (Suggestions for counting approach and how to decide which ones pass the threshold.)
Part 2: Flow cytometry analysis
Save-as Excel sheet, keep just what they need.
Prepare separate sheet with columns/calculations of interest.
Finally, copy in the data.
Add all B/G ratios and NHEJ ratios group Excel worksheet somehow?
For next time
Methods, as promised.
Reagent list
write something here or not accessible to edit
Previous Day: DNA repair assays