Talk:IGEM:MIT/2006/Communications

• Author guidlines for Nature MSB
• PNAS information for authors

Questions for Stephen

1. 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)
   • A: We used the Knight protocol.
     • Then are you sure that you phosphorylated the primers? The Knight protocol doesn't call for this step.

• I'm sure we did.

1. 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.

1. 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

General outstanding issues

• Funding sources for each person

1. See last email... all undergrads were funded in the summer of 2006 by the MIT UROP office

• Protocol for transformation

See below

1. 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>