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| - | {{:Team:Grenoble/Templates/Biology}}
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| - | <html xmlns="http://www.w3.org/1999/xhtml" xml:lang="en" lang="en">
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| - | <body id="Biology">
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| - | <div id="cadre">
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| - | <section>
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| - | <h1>August</h1>
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| - | <a href="https://2012.igem.org/Team:Grenoble/Biology/Notebook/August">Week 31</a> •
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| - | <a href="https://2012.igem.org/Team:Grenoble/Biology/Notebook/August/week_32">Week 32</a> •
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| - | <a href="https://2012.igem.org/Team:Grenoble/Biology/Notebook/August/week_33">Week 33</a> •
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| - | <a href="https://2012.igem.org/Team:Grenoble/Biology/Notebook/August/week_34">Week 34</a> •
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| - | <a href="https://2012.igem.org/Team:Grenoble/Biology/Notebook/August/week_35">Week 35</a>
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| - | </section>
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| - | <br/>
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| - | <section>
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| - | <h1> Week 33: August 13<span class="exposant">th</span> to 19<span class="exposant">th</span> </h1>
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| - | <h2> Goal of the week: </h2>
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| - | Assembly of:
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| - | <ul><li>pAra/Bad_RBS_GFP and RBS_Cya.</li>
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| - | <li>pompC and mcherry</li>
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| - | <li>Tap and EnvZ</li>
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| - | </ul>
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| - | </section>
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| - | <section>
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| - | <h2> Monday, August 13<span class="exposant">th</span>:</h2>
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| - | We did some verification PCRs on miniprep with HF Phusion enzyme (<a href="https://2012.igem.org/Team:Grenoble/Biology/Protocols/PCR_2">protocol</a>) in order to check the
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| - | Gibson Assembly products (12/09/09>.<br/>Annealing temperature = 55°C.<br/>
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| - | <br/>
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| - | To separate (<a href="https://2012.igem.org/Team:Grenoble/Biology/Protocols/Gel">protocol</a>) the PCR products, we prepared a 1.8% TAE agarose gel.<br/>
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| - | Migration conditions = 100V during 30 min.<br/>
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| - | In order to reveal the DNA fragments, we used <a href="https://2012.igem.org/Team:Grenoble/Biology/Protocols/EtBr">EtBr</a>.<br/>
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| - | <br/>
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| - | It showed no significant results (data not shown).
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| - | </section>
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| - |
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| - | <section>
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| - | <h2> Tuesday, August 14<span class="exposant">th</span>:</h2>
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| - | We did an experiment (<a href="https://2012.igem.org/Team:Grenoble/Biology/Protocols/AND_test">protocol</a>) in order to test the "AND" gate (CRP-cAMP & araC) without the amplifier module. <br/>
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| - | <br/>
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| - | We transformed (new <a href="https://2012.igem.org/Team:Grenoble/Biology/Protocols/Transformation_2">protocol</a>) BW25113 WT
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| - | competent cells (<a href="https://2012.igem.org/Team:Grenoble/Biology/Protocols/Competence">protocol</a>) with pSB3C5.<br/>
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| - | </section>
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| - |
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| - | <section>
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| - | <h2> Wednesday, August 15<span class="exposant">th</span>:</h2>
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| - | We did an experiment (<a href="https://2012.igem.org/Team:Grenoble/Biology/Protocols/AND_test">protocol</a>) in order to test the "AND" gate (CRP-cAMP & araC) without the amplifier module. The M9 medium used was complemented: 0.1% acetate.
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| - | </section>
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| - | <section>
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| - | <h2>Thursday, August 16<span class="exposant">th</span>:</h2>
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| - | We did some colony PCRs with HF Phusion enzyme (<a href="https://2012.igem.org/Team:Grenoble/Biology/Protocols/PCR_2">protocol</a>) in order to amplify pSB1A3 (araC), pSB3C5 (araC), pSB3C5 (pAra/Bad_RBS_GFP_RBS_Cya) and pSB4C5 (pAra/Bad_RBS_GFP_RBS_Cya)<br/>
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| - | <br/>
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| - | Then, we did some digestions (<a href="https://2012.igem.org/Team:Grenoble/Biology/Protocols/Restriction">protocol</a>). The digestions were achieved with two restriction enzymes:
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| - | <ul>
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| - | <li> pOmpC with EcoRI and SpeI during 10 minutes </li>
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| - | <li> pOmpC with EcoRI</li>
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| - | <li> pSB4K5 with EcoRI and SpeI during 10 minutes</li>
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| - | <li> pSB4K5 with EcoRI during 10 minutes</li>
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| - | </ul>
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| - | <br/>
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| - | To separate (<a href="https://2012.igem.org/Team:Grenoble/Biology/Protocols/Gel">protocol</a>) the PCR and the digestion products, we prepared a 1.8% TAE agarose gel.<br/>
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| - | Migration conditions = 100V during 30 min.<br/>
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| - | In order to reveal the DNA fragments, we used <a href="https://2012.igem.org/Team:Grenoble/Biology/Protocols/EtBr">EtBr</a>.<br/>
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| - | <br/>
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| - | The PCR and the digestion worked well, we realised a DNA extraction (<a href="https://2012.igem.org/Team:Grenoble/Biology/Protocols/DNA_extract">protocol</a>) from the pSB1A3 (araC) and pSB4C5 (pAra/Bad_RBS_GFP_RBS_Cya) PCR products.<br/>
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| - | <br/>
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| - | </div>
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| - | </body>
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| - | </html>
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| - | {{:Team:Grenoble/script}}
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| - | {{:Team:Grenoble/menu}}
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