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

Line 1: Line 1:
 
{{Template:Paris_Bettencourt}}
 
{{Template:Paris_Bettencourt}}
 
<html>
 
<html>
−
<b>Thursday, Jul. 23</b>
+
<b>13/07</b>
 
<ul>
 
<ul>
−
<li>Understood the project, just have to do it.</li>
+
<li>Received gBlocks RibA, RibD, RibE, RibT25 and RibT48 and amplification oligos from IDT.</li>
 +
 
 +
Dilution in water and PCR amplification, using the following protocol:
 +
<ul>
 +
<li>1 μL gBlock (0.1 to 1ng)
 +
<li>1 μL forward primer (10 μM)
 +
<li>1 μL reverse primer (10 μM)
 +
<li>22 μL DNAse/RNAse free water
 +
<li>25 μL LifeTech MasterMix (2X)
 
</ul>
 
</ul>
  
−
<b>Friday, Jul. 24</b>
+
<b>RibA</b> + <b>o15.001</b> (GCGCCCGAAGACTTATGCAG) + <b>o15.002</b>(GGCCCCGCGCATATGAAG)</br>
 +
<b>RibD</b> + <b>o15.003</b>(CGCTATAGAAGACTTGAGAAGATCTG) + <b>o15.004</b>(GCGCGGCACCACATATGAAG)</br>
 +
<b>RibE</b> + <b>o15.005</b>(CGGCTATAGAAGACTTGCGC) + <b>o15.006</b>(CGCGCCGGCATATGAAGA)</br>
 +
<b>RibT25</b> + <b>o15.007</b>(CCGCGTATAGAAGACTGCTAGA) + <b>o15.008</b>(CAGCAGCATATGAAGACAACCC)</br>
 +
<b>RibT48</b> + <b>o15.009</b>(GCGGTATAGAAGACTGCTAGAGA) + <b>o15.010</b>(CAGCAGCATATGAAGACAACCC)</br>
 +
</br>
 +
For each PCR reaction, a negative control without matrix DNA was prepared.</br>
 +
 
 +
30sec at 98°C</br>
 +
12 cycles</br>
 +
      10sec at 98°C</br>
 +
      30sec at 52°C</br>
 +
      1min at 72°C</br>
 +
</br>
 +
10min at 72°C</br>
 +
until opening at 12°C</br>
 +
</br>
 +
</br><b>Friday, Jul. 24</b>
 
<ul>
 
<ul>
 
<li>Made the project, it is awesome!</li>
 
<li>Made the project, it is awesome!</li>

Revision as of 14:54, 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


    Friday, Jul. 24
    • Made the project, it is awesome!

    KOIN!