Talk:IGEM:MIT/2006/Communications

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(Questions for Stephen)
(General outstanding issues)
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*Funding sources for each person
*Funding sources for each person
*Protocol for transformation
*Protocol for transformation
 +
See below
 +
 +
#Transformation - André [http://openwetware.org/wiki/Transforming_chemically_competent_cells_%28Inoue%29 Transform competent cells]
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#*Thaw ~50 μl cells (whichever you like) on ice.  Do not use glass tubes, which adsorb DNA.
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#*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.
*Describing figure 6
*Describing figure 6
*Update <bbpart>BBa_Q04401</bbpart>
*Update <bbpart>BBa_Q04401</bbpart>

Revision as of 15:48, 11 February 2008


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
  • 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>
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