20.109(S14):Data analysis (Day7): Difference between revisions
From OpenWetWare
Jump to navigationJump to search
Line 16: | Line 16: | ||
===Part 1: Stain irradiated cell colonies=== | ===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=== | ===Part 2: Flow cytometry analysis=== |
Revision as of 09:44, 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
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
For next time
Reagent list
write something here or not accessible to edit
Previous Day: DNA repair assays