Difference between revisions of "Team:TU Dresden/InterLab"

Line 17: Line 17:
 
<h2>Materials and Methods</h2>
 
<h2>Materials and Methods</h2>
 
<p>To generate a reliable and robust standard, certain requirements need to be fulfilled. They include following a precise protocol, the usage of competent <i>Escherichia coli</i> DH5-alpha cells and GFP measurement with a plate reader. For more detailed information, see <a href="https://2017.igem.org/Competition/InterLab_Study">https://2017.igem.org/Competition/InterLab_Study</a></p>
 
<p>To generate a reliable and robust standard, certain requirements need to be fulfilled. They include following a precise protocol, the usage of competent <i>Escherichia coli</i> DH5-alpha cells and GFP measurement with a plate reader. For more detailed information, see <a href="https://2017.igem.org/Competition/InterLab_Study">https://2017.igem.org/Competition/InterLab_Study</a></p>
 +
 +
<h3>Devices</h3>
 +
<p>The following eight devices were used, all composed in the backbone of pSB1C3 including an chloramphenicol resistance gen and a GFP reporter gene (except for the negative control). They differ in the combination of promoter and ribosome binding site (RBS) upstream of the reporter.</p>
 +
 +
<ul>
 +
    <li> Positive Control (<a href="http://parts.igem.org/Part:BBa_I20270">(BBa_I20270)</a>) </li>
 +
    <li> Negative Control (<a href="http://parts.igem.org/Part:BBa_R0040">(BBa_R0040)</a>) </li>
 +
    <li> Test Device 1 (<a href="http://parts.igem.org/Part:BBa_J364000">BBa_J364000</a>) </li>
 +
    <li> Test Device 2 (<a href="http://parts.igem.org/Part:BBa_J364001">BBa_J364001</a>) </li>
 +
    <li> Test Device 3 (<a href="http://parts.igem.org/Part:BBa_J364002">BBa_J364002</a>) </li>
 +
    <li> Test Device 4 (<a href="http://parts.igem.org/Part:BBa_J364003">BBa_J364003</a>) </li>
 +
    <li> Test Device 5 (<a href="http://parts.igem.org/Part:BBa_J364004">BBa_J364004</a>) </li>
 +
    <li> Test Device 6 (<a href="http://parts.igem.org/Part:BBa_J364005">BBa_J364005</a>) </li>
 +
</ul>
 +
 +
<h3>Transformation of plasmids into <i>E. coli</i> DH5α</h3>
 +
<p>In a first step, the given plasmids (provided in the Measurement Kit) had to be transformed into <i>E. coli</i> DH5α cells. We used the provided <a href="http://parts.igem.org/Help:Protocols/Transformation">Transformation protocol</a></p>
  
 
</div>
 
</div>

Revision as of 11:41, 26 October 2017

International InterLab Measurement Study

Background

For achieving reliable results there is a need for repetition of experiments. In this course, maybe the most ambitious aspect is the repetition of measurements in different labs: the usage of different strains, protocols and devices makes it hard to generate comparable results.

By now, fluorescent proteins such as GFP became useful and widely applied tools in synthetic biology. The comparison of fluorescence data from different labs is even more challenging as they might be reported in different ways or generated by various techniques. Therefore, the Measurement committee uses the framework of the international competition iGEM to gain experimental data from all over the world. 2017, the fourth International InterLab Measurement Study was set-up with the aim to develop a robust GFP measurement. We as the TU_Dresden Team also work with fluorescent proteins ourselves, therefore we definitely wanted to participate in the this years InterLab Study.

“How close can the numbers be when fluorescence is measured all around the world?” (iGEM Measurement Committee)

Materials and Methods

To generate a reliable and robust standard, certain requirements need to be fulfilled. They include following a precise protocol, the usage of competent Escherichia coli DH5-alpha cells and GFP measurement with a plate reader. For more detailed information, see https://2017.igem.org/Competition/InterLab_Study

Devices

The following eight devices were used, all composed in the backbone of pSB1C3 including an chloramphenicol resistance gen and a GFP reporter gene (except for the negative control). They differ in the combination of promoter and ribosome binding site (RBS) upstream of the reporter.

Transformation of plasmids into E. coli DH5α

In a first step, the given plasmids (provided in the Measurement Kit) had to be transformed into E. coli DH5α cells. We used the provided Transformation protocol