Team:Evry/Notebook/w12

From 2013.igem.org

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<h1>Week 12: 2nd September - 8th September</h1>
<h1>Week 12: 2nd September - 8th September</h1>
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<img src="http://img.xooimage.com/files10/9/6/9/avatar.02.09.2-de3506.gif" alt="Miam" />
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<h2> Plasmid 3<h/2>
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We launch another PC to get Enterobactins A, B, C, D, E and F genes.
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Add the recquired primers in each tube:
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<ol>
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<li> EntA: primers 065 and 066
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<li> EntB: primers 067 and 068
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<li> EntC: primers 069 and 070
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<li> EntD: primers 071 and 072
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<li> EntE: primers 073 and 074
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<li> EntF: primers 075 and 076
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<li> Positive controle: primers 009 and 010
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<li> Negative controle: primers 009 and 010 </ol> <br/>
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<i>we use pEntC as our controle.<br/>
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For more details about our primers, see the <a href='https://2013.igem.org/Team:Evry/Primers' target='_blank'>corresponding page</a>.<br/>
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We then prepared a master mix 1 with:
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<ul>
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<li> 9 x 27,5 = 247,5 µL of distilled water
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<li> 9 x 1 = 9 µL of dNTPs
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<li> 9 x 10 = 90 µL of Q<inf>5</inf> Buffer
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<li> 9 x 0,5 = 4,5 µL of Q<inf>5</inf>
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For the negative controle, we added 1 µL of distilled water in tube 8 and 39 µL of master mix 1.<br/>
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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.<br/>
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Revision as of 13:02, 2 September 2013

Tex am:Evry/template notebook

Week 12: 2nd September - 8th September

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.

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