Difference between revisions of "Team:DTU-Denmark/Experiments"

Line 31: Line 31:
  
 
<H2 style="text-align: left;margin-bottom:20px;">Synthlab protocols</H2>
 
<H2 style="text-align: left;margin-bottom:20px;">Synthlab protocols</H2>
−
<button class="collapsible">Open Section 1</button>
+
<button class="collapsible">Resuspension of DNA/primer</button>
 
<div class="content">
 
<div class="content">
 +
<div class="row">
 +
<div class="col-xs-12">
 +
 +
<p style="font-size: 70%;" >
 +
DNA from IDT will typically be delivered in a white flaky substance, which need to be resuspended, before the DNA is ready for use
 +
</p>
 +
 +
 +
 +
</div>
 +
<div class="col-xs-4">
 +
<h4 class="media heading">Materials</h4>
 +
 +
<ul>
 +
  <li><p style="font-size:90%">Table centrifuge</p></li>
 +
<li><p style="font-size:90%">EB/TE buffer</p></li>
 +
<li><p style="font-size:90%">Genes/Primers from IDT or other DNA provider</p></li>
 +
</ul>
 +
 +
 +
 +
</div>
 +
<div class="col-xs-8">
 +
 +
<h4 class="media heading">Procedure</h4>
 +
 +
<ol>
 +
  <li><p style="font-size:90%">Quickly spin the DNA down in the table centrifuge</p></li>
 +
<li><p style="font-size:90%">Calculate the amount of EB buffer need to dilute the gblocks to a desired concentration. Important: gene fragments and primeres are not diluted to the same concentration: The concentration of gene fragments is usually 25 ng/μL  and for primers it is 100 μM.</p></li>
 +
<li><p style="font-size:90%">Genes/Primers from IDT or other DNA provider</p>
 +
<ul>
 +
  <li><p style="font-size:90%">DNA from IDT usually comes in dried flakes of 500 or 1000 ng of DNA. To achieve the desired concentration (usually 25 ng/μL) the needed amount of EB buffer is  40 μL (for 1000 ng samples) or 20 μL (for 500 ng samples)</p></li>
 +
<li><p style="font-size:90%"> In order to calculate the the molar amount of primer, use the <a href="https://nebiocalculator.neb.com/#!/ssdnaamt">NEB calculator</a></p></li>
 +
</ul>
 +
</li>
 +
 +
<li><p style="font-size:90%">Add the calculated amount of EB buffer</p></li>
 +
<li><p style="font-size:90%">Store the resuspended DNA at -20°C </p></li>
 +
</ol>
 +
 +
</div>
 +
 +
 +
 +
 
   <p>Lorem ipsum dolor sit amet, consectetur adipisicing elit, sed do eiusmod tempor incididunt ut labore et dolore magna aliqua. Ut enim ad minim veniam, quis nostrud exercitation ullamco laboris nisi ut aliquip ex ea commodo consequat.</p>
 
   <p>Lorem ipsum dolor sit amet, consectetur adipisicing elit, sed do eiusmod tempor incididunt ut labore et dolore magna aliqua. Ut enim ad minim veniam, quis nostrud exercitation ullamco laboris nisi ut aliquip ex ea commodo consequat.</p>
 
</div>
 
</div>

Revision as of 10:12, 14 September 2018

Experiments

Synthlab protocols

DNA from IDT will typically be delivered in a white flaky substance, which need to be resuspended, before the DNA is ready for use

Materials

  • Table centrifuge

  • EB/TE buffer

  • Genes/Primers from IDT or other DNA provider

Procedure

  1. Quickly spin the DNA down in the table centrifuge

  2. Calculate the amount of EB buffer need to dilute the gblocks to a desired concentration. Important: gene fragments and primeres are not diluted to the same concentration: The concentration of gene fragments is usually 25 ng/μL and for primers it is 100 μM.

  3. Genes/Primers from IDT or other DNA provider

    • DNA from IDT usually comes in dried flakes of 500 or 1000 ng of DNA. To achieve the desired concentration (usually 25 ng/μL) the needed amount of EB buffer is 40 μL (for 1000 ng samples) or 20 μL (for 500 ng samples)

    • In order to calculate the the molar amount of primer, use the NEB calculator

  4. Add the calculated amount of EB buffer

  5. Store the resuspended DNA at -20°C

Lorem ipsum dolor sit amet, consectetur adipisicing elit, sed do eiusmod tempor incididunt ut labore et dolore magna aliqua. Ut enim ad minim veniam, quis nostrud exercitation ullamco laboris nisi ut aliquip ex ea commodo consequat.

Lorem ipsum dolor sit amet, consectetur adipisicing elit, sed do eiusmod tempor incididunt ut labore et dolore magna aliqua. Ut enim ad minim veniam, quis nostrud exercitation ullamco laboris nisi ut aliquip ex ea commodo consequat.

Lorem ipsum dolor sit amet, consectetur adipisicing elit, sed do eiusmod tempor incididunt ut labore et dolore magna aliqua. Ut enim ad minim veniam, quis nostrud exercitation ullamco laboris nisi ut aliquip ex ea commodo consequat.