Difference between revisions of "Team:Madrid-OLM/AptamerProtocols"

Line 67: Line 67:
 
                                         <div class="tab__content">
 
                                         <div class="tab__content">
 
                                             <h3>SELEX</h3>
 
                                             <h3>SELEX</h3>
−
                                            <h5>Prepare the library pool</h5>
 
−
                                           
 
 
                                             <ol class="ourlist">
 
                                             <ol class="ourlist">
−
                                                <li class="nomargin"> <p class="lead">Resuspend 2 nmol de la library pool on 200 µl of Binding Buffer (BB).</p></li>
 
−
                                                <li class="nomargin"><p class="lead">Denatured the library by heating it at 90ºC for 10 min and immediately cold it on ice for another 10 min.</p></li>
 
−
                                                <li class="nomargin"><p class="lead">To get rid of the DNA that unespecifically binds to the system, apply the library through  a nitrocellulose  membrane and centrifuge 1 min at 8000 rpm. Quantify the DNA that does  not bind unspeficically and note it as the initial DNA.</p></li>
 
 
                                                 <h4>Prepare the library pool</h4>
 
                                                 <h4>Prepare the library pool</h4>
 
                                                 <li class="nomargin"> <p class="lead">Resuspend 2 nmol de la library pool on 200 µl of Binding Buffer (BB).</p></li>
 
                                                 <li class="nomargin"> <p class="lead">Resuspend 2 nmol de la library pool on 200 µl of Binding Buffer (BB).</p></li>
 
                                                 <li class="nomargin"><p class="lead">Denatured the library by heating it at 90ºC for 10 min and immediately cold it on ice for another 10 min.</p></li>
 
                                                 <li class="nomargin"><p class="lead">Denatured the library by heating it at 90ºC for 10 min and immediately cold it on ice for another 10 min.</p></li>
 
                                                 <li class="nomargin"><p class="lead">To get rid of the DNA that unespecifically binds to the system, apply the library through  a nitrocellulose  membrane and centrifuge 1 min at 8000 rpm. Quantify the DNA that does  not bind unspeficically and note it as the initial DNA.</p></li>
 
                                                 <li class="nomargin"><p class="lead">To get rid of the DNA that unespecifically binds to the system, apply the library through  a nitrocellulose  membrane and centrifuge 1 min at 8000 rpm. Quantify the DNA that does  not bind unspeficically and note it as the initial DNA.</p></li>
 +
                                               
 +
                                                <h4>Protein-Aptamer incubation</h4>
 +
                                                <li class="nomargin"> <p class="lead">Incubate the flowthrough with the protein of interest during 1 hour.</p></li>
 +
                                                <li class="nomargin"><p class="lead">Apply the DNA to a new nitrocellulose  membrane as in step 3.</p></li>
 +
                                                <li class="nomargin"><p class="lead">Wash the membrane four times with 300 µl of BB, like on step 3.</p></li>
 +
                                                <li class="nomargin"><p class="lead">Recover the membrane and transfer it to a new Eppendorf  tube  without letting it dry.  </p></li>
  
  

Revision as of 12:44, 10 October 2018

Madrid-OLM

Aptamer`s Protocols

Aptamer's Protocols

Texto de explicacion/ resumen de la pagina.

Aptamer Discovery

  • SELEX

    SELEX

      Prepare the library pool

    1. Resuspend 2 nmol de la library pool on 200 µl of Binding Buffer (BB).

    2. Denatured the library by heating it at 90ºC for 10 min and immediately cold it on ice for another 10 min.

    3. To get rid of the DNA that unespecifically binds to the system, apply the library through a nitrocellulose membrane and centrifuge 1 min at 8000 rpm. Quantify the DNA that does not bind unspeficically and note it as the initial DNA.

      4. Protein-Aptamer incubation

    4. Incubate the flowthrough with the protein of interest during 1 hour.

    5. Apply the DNA to a new nitrocellulose membrane as in step 3.

    6. Wash the membrane four times with 300 µl of BB, like on step 3.

    7. Recover the membrane and transfer it to a new Eppendorf tube without letting it dry.

  • PCI

    PCI

  • QiaGen

    QiaGen



Back to Dicovery Protocol Index

Aptamer Characterization

Elona