Talk:IGEM:MIT/2006/Communications: Difference between revisions
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#*110 RPM- the shaker by Samantha's/2007 iGEM team's bench | #*110 RPM- the shaker by Samantha's/2007 iGEM team's bench | ||
#Are both J45200 and J45250 on pSB1AT3? | #Are both J45200 and J45250 on pSB1AT3? | ||
#Check with Francois re rpoS promoter plasmid | |||
#Check with Alex and Amy re constant pressure. | |||
==General outstanding issues== | ==General outstanding issues== | ||
Revision as of 14:25, 11 February 2008
Questions for Stephen
- Do we have the regulated wintergreen GC data in a form that we could generate plots similar to the banana timecourse?
- Yes, we could. It would take some digging through old files and sorting out through each protocol we used on each day. I don't know if it's necessary to show though.
- What did you use to shake the cultures? i.e. what incubator or shaker?
- 220 RPM- the shaker in 37 C room
- 110 RPM- the shaker by Samantha's/2007 iGEM team's bench
- Are both J45200 and J45250 on pSB1AT3?
- Check with Francois re rpoS promoter plasmid
- Check with Alex and Amy re constant pressure.
General outstanding issues
- Funding sources for each person
- See last email... all undergrads were funded in the summer of 2006 by the MIT UROP office
- Protocol for transformation
See below
- Transformation - André Transform competent cells
- Thaw ~50 μl cells (whichever you like) on ice. Do not use glass tubes, which adsorb DNA.
- Add 200 uL DNA to cells
- Incubate on ice for 30 minutes
- Incubate cells for 50-60 seconds at 42°C.
- Incubate cells on ice for 2 min.
- Add 200-300μL of room temperature SOC (not critical)
- Incubate for 1 hour at 37°C on shaker.
- Spread 200 μl onto plates made with appropriate antibiotic.
- Grow overnight at 37°C.
- Describing figure 6
- Update <bbpart>BBa_Q04401</bbpart>
