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

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! Green FD buffer (10x)
! Green FD buffer (10x)
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! DNA
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! PstI
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! Xba1  
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Revision as of 11:43, 15 July 2010

iGEM iGarden <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
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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 Digestion Reactions
Mira N1 Mira N2 Mira C1 Mira C2 Brazz N1 Brazz N2 Brazz C1 Brazz C2
diH2O 13.5 13.5 13.5 11 11 11 13.5 13
Green FD buffer (10x) 2 2 2 2 2 2 2 2
DNA 2.5 2.5 2.5 5 5 5 2.5 3
PstI 1 1 1 1 1 1 1 1
Xba1 1 1 1 1 1 1 1 1


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

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

Team Vector


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