- Materials
- LFM
- LFM+1M sorbitol
- Flow Cells
- Ethanol (70%)
- H20
- conA
- Run fluorescene first as a control or do you think we are ok ???
- Methods (general)
- Inoculate cells t = -38 hrs
- Set back cells t = -14 hrs
- Set up flow cell
- plasma clean, holes, etc.
- wash flow cell with Eth and water
- add ConA for at least 5 mins
- measure OD of cells
- add cells and allow to sit for 5 mins
- Measurement Parameters
- Times
- 210
- 180
- 150
- 120
- 100
- 90
- 80
- 60
- 30
- 10
- Methods (specific)
- Set1: 210 and 180
- Set2: 150 and 120
- run Set1Wt, Set1Mutant, Set2Wt, Set2 Mutant on one day
- Set3: 100 and 90 and 80
- Set4: 60 and 30 and 10
- run Set3Wt, Set3Mutant, Set4Wt, Set4Mutant on another day
- Never sleep
- 5 oscillations per time ie for 60 secs run 11 minutes, start and end with off
- Wait 20 minutes to allow cells to reset in between different times of the same set
- I need to go through the timing, but i think i need 3 days to get through each mutant
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