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Team:Trieste/notebook
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We prepared the antibiotics stock ( Amp, Kan, Cm, Spec ) and the DH5-alpha competent cells.</br> | We prepared the antibiotics stock ( Amp, Kan, Cm, Spec ) and the DH5-alpha competent cells.</br> | ||
</br> | </br> | ||
| - | We extracted the following BB from the kit: the constitutive promoter ( | + | We extracted the following BB from the kit: the constitutive promoter (BBa_J23100), the strong RBS (BBa_B0034), the weak RBS (BBa_B0031), the GFP (BBa_E0240) the double terminator (BBa_B0015), the toxin Tse2 (BBa_K314200).</br> |
| - | < | + | <i>E.coli</i> DH5-alpha competent cells were transformed with 2 ul of each BB extracted.</br> |
| - | E.coli DH5-alpha competent cells were transformed with 2 ul of each BB extracted.</br> | + | |
</br> | </br> | ||
We learnt the safety rules and mostly of the lab techniques. | We learnt the safety rules and mostly of the lab techniques. | ||
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<div id="antibody" class="notebook_section"> | <div id="antibody" class="notebook_section"> | ||
<h2 class="notebook_title">Antibody</h2> | <h2 class="notebook_title">Antibody</h2> | ||
| - | + | The first week in the lab we prepared competent <i>E. coli</i> strains DH5-alpha, W3110 and HB2151 necessary for future experiments. Competent W3110 and HB2151 were first transformed with p-REP 4 plasmid which codes for Lac repressor. The recombinant colonies selected on plate containing kanamycin were inoculated the and made them competent.</br></br> | |
| + | We also digested and purified the synthetized genes from pUC57 vector. | ||
</div> | </div> | ||
<div id="chassis" class="notebook_section"> | <div id="chassis" class="notebook_section"> | ||
| - | <h2 class="notebook_title"> | + | <h2 class="notebook_title">Cumate-Switch Regulation</h2> |
| - | + | We waited the synthetic DNA (RBS - CymR - SV5 tag and T5 promoter - Cumate operator). | |
</div> | </div> | ||
</div> | </div> | ||
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<div id="box_right"> <!-- start box_right --> | <div id="box_right"> <!-- start box_right --> | ||
<ul id="sub_menu"> | <ul id="sub_menu"> | ||
| - | <li><a href="https://2012.igem.org/Team:Trieste/notebook">Week 1</a></li> | + | <li class="select" ><a href="https://2012.igem.org/Team:Trieste/notebook">Week 1</a></li> |
<li><a href="https://2012.igem.org/Team:Trieste/notebook2">Week 2</a></li> | <li><a href="https://2012.igem.org/Team:Trieste/notebook2">Week 2</a></li> | ||
<li><a href="https://2012.igem.org/Team:Trieste/notebook3">Week 3</a></li> | <li><a href="https://2012.igem.org/Team:Trieste/notebook3">Week 3</a></li> | ||
| Line 43: | Line 43: | ||
<li><a href="https://2012.igem.org/Team:Trieste/notebook11">Week 11</a></li> | <li><a href="https://2012.igem.org/Team:Trieste/notebook11">Week 11</a></li> | ||
<li><a href="https://2012.igem.org/Team:Trieste/notebook12">Week 12</a></li> | <li><a href="https://2012.igem.org/Team:Trieste/notebook12">Week 12</a></li> | ||
| + | <li><a href="https://2012.igem.org/Team:Trieste/notebooklast">Post European-Jamboree</a></li> | ||
</ul> | </ul> | ||
<img src="https://static.igem.org/mediawiki/2012/b/b0/Team_trieste.jpg" alt="Team iGEM 2012" id="igem_team" /> | <img src="https://static.igem.org/mediawiki/2012/b/b0/Team_trieste.jpg" alt="Team iGEM 2012" id="igem_team" /> | ||
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Follow us also: | Follow us also: | ||
<a href="https://www.facebook.com/IGEMUNITS?ref=ts" target="_blank"><img src="https://static.igem.org/mediawiki/2012/f/f4/Ico_fb.png" alt="Facebook" class="fb" /></a> | <a href="https://www.facebook.com/IGEMUNITS?ref=ts" target="_blank"><img src="https://static.igem.org/mediawiki/2012/f/f4/Ico_fb.png" alt="Facebook" class="fb" /></a> | ||
| - | <a href="https://twitter.com/ | + | <a href="https://twitter.com/iGEMTrieste" target="_blank"><img src="https://static.igem.org/mediawiki/2012/2/21/Ico_twitter.png" alt="twitter" class="tw" /></a> |
</div> | </div> | ||
</div> | </div> | ||
Latest revision as of 18:05, 26 October 2012
The Jolly JoCare
a safe probiotic platform for protein expression
Week 1
More
Suicide System
We prepared the antibiotics stock ( Amp, Kan, Cm, Spec ) and the DH5-alpha competent cells. We extracted the following BB from the kit: the constitutive promoter (BBa_J23100), the strong RBS (BBa_B0034), the weak RBS (BBa_B0031), the GFP (BBa_E0240) the double terminator (BBa_B0015), the toxin Tse2 (BBa_K314200). E.coli DH5-alpha competent cells were transformed with 2 ul of each BB extracted. We learnt the safety rules and mostly of the lab techniques.Antibody
The first week in the lab we prepared competent E. coli strains DH5-alpha, W3110 and HB2151 necessary for future experiments. Competent W3110 and HB2151 were first transformed with p-REP 4 plasmid which codes for Lac repressor. The recombinant colonies selected on plate containing kanamycin were inoculated the and made them competent. We also digested and purified the synthetized genes from pUC57 vector.Cumate-Switch Regulation
We waited the synthetic DNA (RBS - CymR - SV5 tag and T5 promoter - Cumate operator).
