IGEM:Harvard/2009/Notebook/Harvard iGEM 2010/2010/07/15: Difference between revisions

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==Team Flavor==
==Team Flavor==
===Miraculin/Brazzein YFP N/C and NosT + Stop in V24 DUET vector===
* Made glycerol stocks (666 μL glycerol and 333 μL cells)
    ODs
    Miraculin C1: 168.8 ng/μL
    Miraculin C2: 67.2 ng/μL
    Miraculin N1: 183.0 ng/μL
    Miraculin N2: 173.5 ng/μL
    Brazzein N1: 54.8 ng/μL
    Brazzein N2: 79.0 ng/μL
    Brazzein C1: 161.4 ng/μL
    Brazzein C2: 125.0 ng/μL


''Confirmation Digest of Mira/Brazz + YFP + NOSt + STOP in V24 Expression Vector'' <br/>
''Confirmation Digest of Mira/Brazz + YFP + NOSt + STOP in V24 Expression Vector'' <br/>

Revision as of 11:36, 15 July 2010

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Team Allergy

  • Minipreps of PDK+V0120 and GerS+PDK (LTPS+ PDK did not grow)
    • glycerol stocks 500uL of 80% glycerol + 500 uL cells
    • Redoing Ligations/Transformations of LTPS+PDK
    • Sending PDK+V0120 for sequencing
  • Looking at sequencing results for our other amiRNA parts
    • Bet did not work
    • GFP worked
  • Minipreps of Bet 1S (2); Bet 1.2 S (1); Bet 1.2 A (1)
    • Repicking colonies for Ger S/A, LTPA, BetA,
  • Looking at PCR results for Allergen Panel from Genomic DNA that we extracted (did not work)
  • Ladder, GFPS, GFPA (These first two should not work), LTPS, LTPA, BetS, BetA, Bet2S, Bet2A, GerS, GerA, PME, PAL, PDK

[[Image: |240px|allergen panel from arabidopsis DNA extraction]]

  • Digest and Ligate into V0120 and Transform
    • Tmrw: grow up cultures for miniprepping and sending out for sequencing

Team Fence

Team Flavor

Miraculin/Brazzein YFP N/C and NosT + Stop in V24 DUET vector

  • Made glycerol stocks (666 μL glycerol and 333 μL cells)
   ODs
    Miraculin C1: 168.8 ng/μL
    Miraculin C2: 67.2 ng/μL
    Miraculin N1: 183.0 ng/μL
    Miraculin N2: 173.5 ng/μL
    Brazzein N1: 54.8 ng/μL
    Brazzein N2: 79.0 ng/μL
    Brazzein C1: 161.4 ng/μL
    Brazzein C2: 125.0 ng/μL


Confirmation Digest of Mira/Brazz + YFP + NOSt + STOP in V24 Expression Vector 

   Expected Lengths
    Miraculin + YFP + NOSt + STOP = 700 + 1500 + 250 = 2450 bp
    Brazzein + YFP + NOSt + STOP = 300 + 1500 + 250 = 2050 bp
    V24 = 5200 bp


  1. Ladder
  2. Mira N1
  3. Mira N2
  4. Mira C1
  5. Mira C2
  6. Brazz N1
  7. Brazz N2
  8. Brazz C1
  9. Brazz C2
  10. Ladder
  • The N & C designation corresponds to the terminus upon which the YFP was fused.

Team Vector


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