Difference between revisions of "Team:Paris Bettencourt/Notebook/VitaminB2"

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until opening at 12°C</br>
 
until opening at 12°C</br>
 
</br>
 
</br>
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</br><b>Friday, Jul. 24</b>
 
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<ul>
 
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<li>Made the project, it is awesome!</li>
 
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</ul>
 
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<h2>KOIN!</h2>
 

Revision as of 14:55, 10 August 2015

13/07
  • Received gBlocks RibA, RibD, RibE, RibT25 and RibT48 and amplification oligos from IDT.
    • Dilution in water and PCR amplification, using the following protocol:
    • 1 μL gBlock (0.1 to 1ng)
    • 1 μL forward primer (10 μM)
    • 1 μL reverse primer (10 μM)
    • 22 μL DNAse/RNAse free water
    • 25 μL LifeTech MasterMix (2X)
    RibA + o15.001 (GCGCCCGAAGACTTATGCAG) + o15.002(GGCCCCGCGCATATGAAG)
    RibD + o15.003(CGCTATAGAAGACTTGAGAAGATCTG) + o15.004(GCGCGGCACCACATATGAAG)
    RibE + o15.005(CGGCTATAGAAGACTTGCGC) + o15.006(CGCGCCGGCATATGAAGA)
    RibT25 + o15.007(CCGCGTATAGAAGACTGCTAGA) + o15.008(CAGCAGCATATGAAGACAACCC)
    RibT48 + o15.009(GCGGTATAGAAGACTGCTAGAGA) + o15.010(CAGCAGCATATGAAGACAACCC)

    For each PCR reaction, a negative control without matrix DNA was prepared.
    30sec at 98°C
    12 cycles
    10sec at 98°C
    30sec at 52°C
    1min at 72°C

    10min at 72°C
    until opening at 12°C