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| </p> | | </p> |
| <p style="text-align: justify; margin-bottom: 1em;"> | | <p style="text-align: justify; margin-bottom: 1em;"> |
− | Because we wanted to establish <i>Synechococcus elongatus</i> as a new chassis for the iGEM community and scientists, we should
| + | |
− | find the best conditions for cultivation and the best measuring method for our parts in UTEX 2973. Therefore
| + | |
− | we analyzed a big variety of cultivating conditions in measuring growth curves, tried to find a standard in
| + | |
− | light measurement, evaluated different reporters, established a measurement method and compared it to a
| + | |
− | already known FACS measurement method.
| + | |
− | </p>
| + | |
− | <p style="text-align: justify; margin-bottom: 1em;">
| + | |
− | At the beginning of our project we faced the first question: how to cultivate UTEX at 1500 μE?
| + | |
− | To answer this we had to measure the light conditions in our incubators and while doing this simple task the first
| + | |
− | part of standardization began. We discovered that nearly every paper is using different methods to measure
| + | |
− | their light conditions and that it is a really complex and important procedure. So we got in contact with
| + | |
− | Cyano and light <a href="https://2019.igem.org/Team:Marburg/Human_Practices">measurement experts</a> to confront this problem and standardize it. In the following
| + | |
− | popups we show different ways of measurement, their (dis-)advantages and different results depending on the
| + | |
− | measuring instrument.
| + | |
− | </p>
| + | |
− | <p style="text-align: justify; margin-bottom: 1em;">
| + | |
− | Moreover, not only the light intensity but also a variety of other cultivating parameters needed to be analyzed.
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− | In literature and while talking with different experts, we recognized that small deviations of these
| + | |
− | parameters had a huge impact on the growth speed of <i>Synechococcus elongatus</i>. While establishing UTEX 2973 as
| + | |
− | a new chassis we evaluated this impact on the growth speed and were able to show combinations of parameters
| + | |
− | that lead to the fastest growth speed.
| + | |
− | </p>
| + | |
− | <p style="text-align: justify; margin-bottom: 1em;">
| + | |
− | Other aspects were measuring the expression and characterizing our part. Different possibilities were
| + | |
− | discussed and after testing them we decided on two methods in our project. One
| + | |
− | approach was to measure the fluorescence/luminescence with a plate reader. Plate
| + | |
− | readers belong to standard equipment of every lab nowadays, and could deliver easy reproducible results.
| + | |
− | The second way was to measure the fluorescence by FACS (Fluorescence-Activated Cell Sorting). In
| + | |
− | contrast to a platerader a FACS device delivers results with high accuracy by measuring every cell by its
| + | |
− | own.
| + | |
− | </p>
| + | |
− | <p style="text-align: justify; margin-bottom: 1em;">
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− | However, not
| + | |
− | every laboratory possesses a FACS device. So in the end we would like to offer a database - analyzed using these two methods -
| + | |
− | of our constructs for iGEM teams and research groups, who do not have access to a FACS device and show the
| + | |
− | difference in measurement methods.<br>
| + | |
− | <p style="text-align: justify; margin-bottom: 1em;">
| + | |
− | | + | |
− | In the end of the project we were able to create a protocol how to handle <i>Synechococcus elongatus</i> UTEX 2973
| + | |
− | and make a contribution to the cyano community by establishing essential/fixed standards in measurements.
| + | |
− | </p>
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| </article> | | </article> |
| </section> | | </section> |
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| <div class="popup-container"> | | <div class="popup-container"> |
| <div class="popup-header"> | | <div class="popup-header"> |
− | <h1 class="title">Light Measurement</h1> | + | <h1 class="title">Light measurement</h1> |
| <button type="button" onclick="hide('light_measurement')">X</button> | | <button type="button" onclick="hide('light_measurement')">X</button> |
| </div> | | </div> |