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| *[http://mts-msb.nature.com/cgi-bin/main.plex?form_type=display_auth_instructions Author guidlines for Nature MSB] | | *[http://mts-msb.nature.com/cgi-bin/main.plex?form_type=display_auth_instructions Author guidlines for Nature MSB] |
| *[http://www.pnas.org/misc/iforc.shtml PNAS information for authors] | | *[http://www.pnas.org/misc/iforc.shtml PNAS information for authors] |
| | *[http://www.pnas.org/misc/LaTex.shtml Latex template] |
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| | ==General outstanding issues== |
| | *Make sure J45900 gets into registry |
| | *Submit sequences to Genbank |
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| ==Questions for Stephen== | | ===Odor thresholds=== |
| #Did you use the Knight or Endy site directed mutagenesis protocol? (http://openwetware.org/wiki/IGEM:MIT/2006/Notebook/2006-8-10#Mutagenesis says Knight but you told me Endy)
| | "The odor threshold of a compound is the lowest concentration at which its smell can be detected." |
| #*A: We used the Knight protocol.
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| #**Then are you sure that you phosphorylated the primers? The Knight protocol doesn't call for this step.
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| *I'm sure we did.
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| #Do we have the regulated wintergreen GC data in a form that we could generate plots similar to the banana timecourse?
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| *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.
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| #What did you use to shake the cultures? i.e. what incubator or shaker?
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| 220 RPM- the shaker in 37 C room
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| 110 RPM- the shaker by Samantha's/2007 iGEM team's bench
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| ==General outstanding issues==
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| *Funding sources for each person
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| #See last email... all undergrads were funded in the summer of 2006 by the MIT UROP office
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| *Protocol for transformation
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| See below
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| #Transformation - André [http://openwetware.org/wiki/Transforming_chemically_competent_cells_%28Inoue%29 Transform competent cells]
| | *[http://people.ok.ubc.ca/neggers/Chem422A/The%20Odor%20and%20Aroma%20of%20Wine.pdf Isoamyl acetate] |
| #*Thaw ~50 μl cells (whichever you like) on ice. Do not use glass tubes, which adsorb DNA.
| | *[http://www.sciencelab.com/xMSDS-Methyl_salicylate-9927362 Methyl salicylate] |
| #*Add 200 uL DNA to cells
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| #*Incubate on ice for 30 minutes
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| #*Incubate cells for 50-60 seconds at 42°C.
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| #*Incubate cells on ice for 2 min.
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| #*Add 200-300μL of room temperature [[SOC]] (not critical)
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| #*Incubate for 1 hour at 37°C on shaker.
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| #*Spread 200 μl onto plates made with appropriate antibiotic.
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| #*Grow overnight at 37°C.
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| *Describing figure 6
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| *Update <bbpart>BBa_Q04401</bbpart>
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General outstanding issues
- Make sure J45900 gets into registry
- Submit sequences to Genbank
Odor thresholds
"The odor threshold of a compound is the lowest concentration at which its smell can be detected."
