Team:CIDEB-UANL Mexico/Test

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     <meta name="description" content="Cideb Igem Wiki" />
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    <title> Web Site for Igem project of CIDEB </title>
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<title> Web Site for Igem project of CIDEB </title>
      
      
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       <h1> Welcome to the wiki of iGEM CIDEB-UANL_Mexico </h1>
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       <h1> Wet lab </h1>
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           <p>The detection of various components in a safe and rapid way has been a challenge in modern science. Biosensors using genetic circuits in bacteria have been made, which allow knowing whether the sensed component is present or not. Nowadays it is known that certain levels of heavy metals on water can be dangerous for living organisms. For that reason it is important to know the concentration of these metals, but the techniques for detection and quantification are complex and require expensive equipment.</p>
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           <p>In the laboratory we wee working in the construction of the circuit. In the very beggining we didn't have no equipment to use in molecular biology. The principal helped us with the equipment required to work in the laboratory and we started working in March. In a general way here are explained the steps we followed. </p>
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             <p>The aim of this project is to make a study model for quantifying heavy metals with fluorescent colors depending on their levels of concentration. In order to do so, a biosensor based on three different fluorescent reporters will be built. Additionally, this design could be improved in order to achieve higher sensitivity by adding more modules to the circuit.</p>
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          <ol><li>The  first step is to hydrate all the biobricks of the list.</li>
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            <p>This genetic circuit could be applied for building biosensors to detect the presence of heavy metals and semi-quantify them. This design will recognize the component that it is meant to be analyzed and give an approximation of the quantity. According to that, it can be useful to know if the concentration is toxic for living organisms.</p>
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            <li>Then  they must be transformed into bacteria. </li>
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<img class= "image-frame" src="http://i.imgur.com/gbud0.jpg" width="280" height="180" />
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            <li>After  that, we add the bacteria into Petri dishes with agar-based growth medium. </li>
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          <img class= "image-frame" src="http://i.imgur.com/NzeFT.jpg" width="280" height="180" />
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             <li>The next step was to resemble in test tubes. </li>
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        <p>Back in the early nineties (yeah, i'm old) i was tripping ballz on acid one night with some friends. At some point one of us got the brilliant idea to test out our clairvoyant abilities under the influence and we set up a nifty experiment where one of us would take a random card out of a playing deck and would try to 'send' the card telepathically to one of the others. When me and my best friend at the time were up, I ended up calling the exact card three times in a row. Pretty much left the room speechless. The weird thing is, when we talked about it later on we both sort of knew beforehand we could do this and couldn't stop smiling during the whole ordeal.</p>
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            <li>Do  MiniPreparation of plasmid DNA with all the biobricks</li>
 +
            <li>Check  the quality of the DNAs by running </li>
 +
          </ol>
 +
          <p>Then the biobricks will follow a different process divided by sections: Stand-by, High concentration, Low  concentration<img class= "image-frame" src="https://static.igem.org/mediawiki/2012hs/8/8f/Stand-bt.png" width="280" height="300" /></p>
 +
          <p> In order to build the section of Stand-by the following steps must be followed: </p>
 +
       
 +
          <ol>
 +
            <li>Cut  the pieces 1-6M with EcoRI and SpeI, and the part 3-12M with XbaI and PstI.</li>
 +
            <li>Ligate  them into the plasmid pSB1T3 (1-7A).</li>
 +
          </ol>
 +
          <p>&nbsp;</p>
 +
          <p><img class= "image-frame" src="https://static.igem.org/mediawiki/2012hs/1/1f/Alta_concentracion.png" width="350" height="300" /></p>
 +
          <p> In order to build the section of High concentration the following  steps must be followed: </p>
 +
          <ol>
 +
          <ol>
 +
            <li> Cut the part 1-6E with EcoRI and SpeI, and the part 1-10E with XbaI  and PstI.</li>
 +
            <li>Ligate  them into the plasmid PBB1K3 (1-5A) and we obtain the part pBAD-RBS</li>
 +
            <li>Cut  the part 1-14N with EcoRI and SpeI , and the part 1-2M </li>
 +
            <li>Ligate  them into the plasmid PSB1T3 (1-7A) and we obtain the part YFP-cI434.</li>
 +
            <li>Cut  the part pBAD-RBS with EcoRI and SpeI and the part YFP-cI434 with XbaI and  PstI.</li>
 +
            <li>Ligate  them into the the plasmid PSB1A3 (1-1G).</li>
 +
          </ol>
 +
          <p><img class= "image-frame" src="https://static.igem.org/mediawiki/2012hs/d/dd/Low_concentration.png" width="450" height="300" /></p>
 +
          <p> In order to build the section of High concentration the following  steps must be followed: </p>
 +
          <ol>
 +
          <ol>
 +
            <li> Cut the part 2-11J with EcoRI and SpeI, and the part 3-12O with XbaI  and PstI.</li>
 +
            <li>Ligate  them into the plasmid PSB1T3 (1-7A) and we obtain the part PRM-RFP.</li>
 +
            <li> Cut the part 2-14A with EcoRI and SpeI, and the part 1-10C with XbaI  and PstI.</li>
 +
            <li>Ligate  them into the plasmid PSB1K3 (1-5A) and we obtain the part pLL-cI.</li>
 +
            <li>Cut  the part 2-12G with EcoRI and SpeI, and the part 1-10G with XbaI and PstI.</li>
 +
            <li>Ligate  them into the plasmid PSB1K3 (1-5A) and we obtain the part phiR73-c22.</li>
 +
            <li>Cut  the part pLL-cI with EcoRI and SpeI, and the part pRM-RFPwith XbaI and PstI.</li>
 +
            <li>Ligate  them into the plasmid PSB1A3 (1-1G) and we obtain the part pLL-cI-pRM-RFP.</li>
 +
            <li>Cut  the part 1-14N with EcoRI and SpeI, and the part phiR73-c22 with XbaI and PstI.</li>
 +
            <li>Ligate  them into the plasmid PSB1T3 (1-7A) and we obtain the part pBad-phiR73-c22.</li>
 +
            <li>Cut  the part pBad-phiR73-c22 with EcoRI and SpeI, and the part pLL-cI-pRM-RFP with  XbaI and PstI.</li>
 +
            <li>Ligate  them into the plasmid PSB1K3 (1-5A) and we obtain the low concentration circuit.</li>
 +
          </ol>
 +
          <p>&nbsp;</p>
 +
          <p>Until now </p>
 +
          <ol>
 +
          <ul>
 +
            <li>We  had some problems transforming the part 1-14N. We ran out of DNA of this  biobrick so we contacted with the IGEM team from the Tecnológico de Monterrey  and they gave us some DNA. But it didn’t work when we try to transform it.</li>
 +
            <li></li>
 +
            <li>We  have DNA in tubes stored and others in the refrigerator of the lab to use them  continuously.</li>
 +
            <li></li>
 +
            <li>We  have cut all the biobricks but after certain time they tend to degradate so we have to do it continuously.</li>
 +
            <li></li>
 +
            <li>The  Ligation 3 is already done. This is composed by the parts: 2-12G and 1-10G.</li>
 +
          </ul>
 +
          <p>&nbsp;</p>
       </div>
       </div>
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<p class="date"> March 25, 2012</p>
<p class="date"> March 25, 2012</p>
<h3>Headline</h3>
<h3>Headline</h3>
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<p>Back in the early nineties (yeah, i'm old) i was tripping ballz on acid one night with some friends. </p>
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<p>blahblahblah </p>
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<p> class="date"> March 26, 2012</p>
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<p class="date"> March 26, 2012</p>
<h3>Headline II</h3>
<h3>Headline II</h3>
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<p>At some point one of us got the brilliant idea to test out our clairvoyant abilities under the influence and we set up a nifty experiment where one of us would take a random card out of a playing deck and would try to 'send' the card telepathically to one of the others. When me and my best friend at the time were up, I ended up calling the exact card three times in a row. </p>
+
<p>blahblahblah </p>
<p class="date"> March 27, 2012</p>
<p class="date"> March 27, 2012</p>
<h3>Headline III</h3>
<h3>Headline III</h3>
-
<p> Pretty much left the room speechless. The weird thing is, when we talked about it later on we both sort of knew beforehand we could do this and couldn't stop smiling during the whole ordeal.</p>
+
<p> blahblahblah</p>
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      </div>
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      </div>
       <div id="footer1">
       <div id="footer1">
       <p class="footer-text"> iGEM High School Division, 2012 edition - CIDEB-UANL_Mexico team </p>
       <p class="footer-text"> iGEM High School Division, 2012 edition - CIDEB-UANL_Mexico team </p>
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        </div>
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      </div>
          
          
  </div>
  </div>

Latest revision as of 17:23, 16 June 2012

Web Site for Igem project of CIDEB

Wet lab

In the laboratory we wee working in the construction of the circuit. In the very beggining we didn't have no equipment to use in molecular biology. The principal helped us with the equipment required to work in the laboratory and we started working in March. In a general way here are explained the steps we followed.

  1. The first step is to hydrate all the biobricks of the list.
  2. Then they must be transformed into bacteria.
  3. After that, we add the bacteria into Petri dishes with agar-based growth medium.
  4. The next step was to resemble in test tubes.
  5. Do MiniPreparation of plasmid DNA with all the biobricks
  6. Check the quality of the DNAs by running

Then the biobricks will follow a different process divided by sections: Stand-by, High concentration, Low concentration

In order to build the section of Stand-by the following steps must be followed:

  1. Cut the pieces 1-6M with EcoRI and SpeI, and the part 3-12M with XbaI and PstI.
  2. Ligate them into the plasmid pSB1T3 (1-7A).

 

In order to build the section of High concentration the following steps must be followed:

    1. Cut the part 1-6E with EcoRI and SpeI, and the part 1-10E with XbaI and PstI.
    2. Ligate them into the plasmid PBB1K3 (1-5A) and we obtain the part pBAD-RBS
    3. Cut the part 1-14N with EcoRI and SpeI , and the part 1-2M
    4. Ligate them into the plasmid PSB1T3 (1-7A) and we obtain the part YFP-cI434.
    5. Cut the part pBAD-RBS with EcoRI and SpeI and the part YFP-cI434 with XbaI and PstI.
    6. Ligate them into the the plasmid PSB1A3 (1-1G).

    In order to build the section of High concentration the following steps must be followed:

      1. Cut the part 2-11J with EcoRI and SpeI, and the part 3-12O with XbaI and PstI.
      2. Ligate them into the plasmid PSB1T3 (1-7A) and we obtain the part PRM-RFP.
      3. Cut the part 2-14A with EcoRI and SpeI, and the part 1-10C with XbaI and PstI.
      4. Ligate them into the plasmid PSB1K3 (1-5A) and we obtain the part pLL-cI.
      5. Cut the part 2-12G with EcoRI and SpeI, and the part 1-10G with XbaI and PstI.
      6. Ligate them into the plasmid PSB1K3 (1-5A) and we obtain the part phiR73-c22.
      7. Cut the part pLL-cI with EcoRI and SpeI, and the part pRM-RFPwith XbaI and PstI.
      8. Ligate them into the plasmid PSB1A3 (1-1G) and we obtain the part pLL-cI-pRM-RFP.
      9. Cut the part 1-14N with EcoRI and SpeI, and the part phiR73-c22 with XbaI and PstI.
      10. Ligate them into the plasmid PSB1T3 (1-7A) and we obtain the part pBad-phiR73-c22.
      11. Cut the part pBad-phiR73-c22 with EcoRI and SpeI, and the part pLL-cI-pRM-RFP with XbaI and PstI.
      12. Ligate them into the plasmid PSB1K3 (1-5A) and we obtain the low concentration circuit.

       

      Until now

        • We had some problems transforming the part 1-14N. We ran out of DNA of this biobrick so we contacted with the IGEM team from the Tecnológico de Monterrey and they gave us some DNA. But it didn’t work when we try to transform it.
        • We have DNA in tubes stored and others in the refrigerator of the lab to use them continuously.
        • We have cut all the biobricks but after certain time they tend to degradate so we have to do it continuously.
        • The Ligation 3 is already done. This is composed by the parts: 2-12G and 1-10G.

         

Latest News

March 25, 2012

Headline

blahblahblah

March 26, 2012

Headline II

blahblahblah

March 27, 2012

Headline III

blahblahblah

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