Team:Grenoble/Biology/Notebook/July/week 33

From 2012.igem.org

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<h1>August</h1>
<h1>August</h1>
<a href="http://2012.igem.org/Team:Grenoble/Biology/Notebook/August">Week 31</a> •  
<a href="http://2012.igem.org/Team:Grenoble/Biology/Notebook/August">Week 31</a> •  
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<a href="http://2012.igem.org/Team:Grenoble/Biology/Notebook/July/week_32">Week 32</a> •  
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<a href="http://2012.igem.org/Team:Grenoble/Biology/Notebook/August/week_32">Week 32</a> •  
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<a href="http://2012.igem.org/Team:Grenoble/Biology/Notebook/July/week_33">Week 33</a> •  
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<a href="http://2012.igem.org/Team:Grenoble/Biology/Notebook/August/week_33">Week 33</a> •  
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<a href="http://2012.igem.org/Team:Grenoble/Biology/Notebook/July/week_34">Week 34</a> •  
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<a href="http://2012.igem.org/Team:Grenoble/Biology/Notebook/August/week_34">Week 34</a> •  
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<a href="http://2012.igem.org/Team:Grenoble/Biology/Notebook/July/week_35">Week 35</a>
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<a href="http://2012.igem.org/Team:Grenoble/Biology/Notebook/August/week_35">Week 35</a>
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Revision as of 22:54, 26 September 2012

iGEM Grenoble 2012

Project

August

Week 31Week 32Week 33Week 34Week 35

Week 33: August 13th to 19th

Goal of the week:

Assembly of:
  • pAra/Bad_RBS_GFP and RBS_Cya.
  • pompC and mcherry
  • Tap and EnvZ

Monday, August 13th:

We did some verification PCRs on miniprep with HF Phusion enzyme (protocol) in order to check the Gibson Assembly products (12/09/09>.
Annealing temperature = 55°C.

To separate (protocol) the PCR products, we prepared a 1.8% TAE agarose gel.
Migration conditions = 100V during 30 min.
In order to reveal the DNA fragments, we used EtBr.

It showed no significant results (data not shown).

Tuesday, August 14th:

We did an experiment (protocol) in order to test the "AND" gate (CRP-cAMP & araC) without the amplifier module.

We transformed (new protocol) BW25113 WT competent cells (protocol) with pSB3C5.

Wednesday, August 15th:

We did an experiment (protocol) in order to test the "AND" gate (CRP-cAMP & araC) without the amplifier module. The M9 medium used was complemented: 0.1% acetate.

Thursday, August 16th:

We did some colony PCRs with HF Phusion enzyme (protocol) in order to amplify pSB1A3 (araC), pSB3C5 (araC), pSB3C5 (pAra/Bad_RBS_GFP_RBS_Cya) and pSB4C5 (pAra/Bad_RBS_GFP_RBS_Cya)

Then, we did some digestions (protocol). The digestions were achieved with two restriction enzymes:
  • pOmpC with EcoRI and SpeI during 10 minutes
  • pOmpC with EcoRI
  • pSB4K5 with EcoRI and SpeI during 10 minutes
  • pSB4K5 with EcoRI during 10 minutes

To separate (protocol) the PCR and the digestion products, we prepared a 1.8% TAE agarose gel.
Migration conditions = 100V during 30 min.
In order to reveal the DNA fragments, we used EtBr.

The PCR and the digestion worked well, we realised a DNA extraction (protocol) from the pSB1A3 (araC) and pSB4C5 (pAra/Bad_RBS_GFP_RBS_Cya) PCR products.