Team:Evry/Notebook/w12

From 2013.igem.org

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For the purpose of a
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<b>03/09/13</b>
<b>03/09/13</b>

Revision as of 15:48, 3 September 2013

Iron coli project

Week 12: 2nd September - 8th September

TECAN

02/09/13
For the purpose of a

03/09/13

Plasmid 3

We launch another PC to get Enterobactins A, B, C, D, E and F genes. Add the recquired primers in each tube:
  1. EntA: primers 065 and 066
  2. EntB: primers 067 and 068
  3. EntC: primers 069 and 070
  4. EntD: primers 071 and 072
  5. EntE: primers 073 and 074
  6. EntF: primers 075 and 076
  7. Positive controle: primers 009 and 010
  8. Negative controle: primers 009 and 010
We use pEntC as our controle.
For more details about our primers, see the corresponding page.
We then prepared a master mix 1 with:
  • 9 x 27,5 = 247,5 µL of distilled water
  • 9 x 1 = 9 µL of dNTPs
  • 9 x 10 = 90 µL of Q5 Buffer
  • 9 x 0,5 = 4,5 µL of Q5 For the negative controle, we added 1 µL of distilled water in tube 8 and 39 µL of master mix 1.
    For other tubes (6 genes and positive controle), we added 8 µL of genomic DNA in the master mix 1. Then add 40 µL of master mix 1' in each tube.