|
|
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| Vielleicht noch ein allgemeinem abstract zu Messung (vergleiche andere WIKIS) | | Vielleicht noch ein allgemeinem abstract zu Messung (vergleiche andere WIKIS) |
| </p> | | </p> |
| + | |
| + | |
| + | |
| + | |
| + | |
| + | |
| + | |
| + | |
| + | |
| + | |
| + | |
| </section> | | </section> |
| <section class="section"> | | <section class="section"> |
Line 106: |
Line 117: |
| microbiology and necessary to compare results with other scientists and reproduce their data. | | microbiology and necessary to compare results with other scientists and reproduce their data. |
| </p> | | </p> |
| + | |
| + | |
| + | |
| + | |
| + | <figure style="float:right; margin-left: 25px;"> |
| + | <img style="height: 719px; width: 1001px;" |
| + | src="https://2019.igem.org/File:T--Marburg--Metabolic--MEP-pathway.png" |
| + | alt="asas"> |
| + | <figcaption style="max-width: 550px; text-align: center"> |
| + | Fig.1 - Comparison of NanoLuc and teLuc Luminescence Spectra in comparison with Synechococcus elongatus |
| + | UTEX |
| + | 2973 Absorption spectra. |
| + | </figcaption> |
| + | </figure> |
| + | |
| + | <p style="text-align: justify; margin-bottom: 1em;"> |
| + | While we wanted to establish Syn. elong. as a new chassis for the iGEM community and scientists we wanted to |
| + | show 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 on how to cultivate UTEX at 1500 μE. [quelle]. |
| + | So 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 measurement experts [link IHP] to confront this problem and standardize it. In the following |
| + | popup we show different ways of measurement, their (dis-)advantages and different results depending on the |
| + | measuring instrument.<br> |
| + | Not only the light intensity but also a variety of other cultivating parameters needed to be analyzed. |
| + | In literature and while talking with different experts (IHP), we recognized that small deviations of these |
| + | parameters had a huge impact on the growth speed of Synechococcus elongatus. 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.<br> |
| + | Another aspect was measuring the expression and characterize our part. Different possibilities were |
| + | discussed and after testing them we decided on two methods in our project (plate reader and FACs). One |
| + | approach was to measure the fluorescence/luminescence with a plate reader [link part measurement]. Plate |
| + | readers belong to standard equipment of every lab nowadays, and could deliver easy reproducible results.<br> |
| + | The second way was to measure the fluorescence by FACS (Fluorescence-Activated Cell Sorting) [link facs]. In |
| + | contrast to a platerader a FACs device delivers results with high accuracy by measuring every cell by its |
| + | own(vielleicht erst spaeter FACS genau erklaeren aber nicht im abtract?). On the other side not |
| + | every laboratory posses a FACs/device. So in the end we would like to offer a two method analyzed database |
| + | from our crontructs for iGEM teams and research groups, who do not have access to a FACS and show the |
| + | difference in measurement methods.<br> |
| + | At the end of the project we were able to create a protocol how to handle Synechococcus elongatus UTEX 2973 |
| + | and make a contribution to the cyano community by establishing essential/fixed standards in measurement. |
| + | </p> |
| + | </article> |
| + | </section> |
| + | <hr> |
| + | <section class="section grid"> |
| + | <div class="sub" |
| + | onclick="popup('rbn1')"> |
| + | <div class="sub-header"> |
| + | <h1> |
| + | L I G H T<br> |
| + | M E A S U R E M E N T |
| + | </h1> |
| + | <hr> |
| + | </div> |
| + | <div class="sub-content"> |
| + | <p> |
| + | Hier bitte den für diese Stelle zutreffenden Text einfügen, wenn dieser fertig ist. |
| + | </p> |
| + | </div> |
| + | </div> |
| <div class="wrap-collabsible" | | <div class="wrap-collabsible" |
| style="margin-bottom: 25px;"> | | style="margin-bottom: 25px;"> |
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| | | |
| | | |
− | <figure style="float:right; margin-left: 25px;">
| |
− | <img style="height: 719px; width: 1001px;"
| |
− | src="https://2019.igem.org/File:T--Marburg--Metabolic--MEP-pathway.png"
| |
− | alt="asas">
| |
− | <figcaption style="max-width: 550px; text-align: center">
| |
− | Fig.1 - Comparison of NanoLuc and teLuc Luminescence Spectra in comparison with Synechococcus elongatus
| |
− | UTEX
| |
− | 2973 Absorption spectra.
| |
− | </figcaption>
| |
− | </figure>
| |
| | | |
− | <p style="text-align: justify; margin-bottom: 1em;">
| |
− | While we wanted to establish Syn. elong. as a new chassis for the iGEM community and scientists we wanted to
| |
− | show 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 on how to cultivate UTEX at 1500 μE. [quelle].
| |
− | So 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 measurement experts [link IHP] to confront this problem and standardize it. In the following
| |
− | popup we show different ways of measurement, their (dis-)advantages and different results depending on the
| |
− | measuring instrument.<br>
| |
− | Not only the light intensity but also a variety of other cultivating parameters needed to be analyzed.
| |
− | In literature and while talking with different experts (IHP), we recognized that small deviations of these
| |
− | parameters had a huge impact on the growth speed of Synechococcus elongatus. 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.<br>
| |
− | Another aspect was measuring the expression and characterize our part. Different possibilities were
| |
− | discussed and after testing them we decided on two methods in our project (plate reader and FACs). One
| |
− | approach was to measure the fluorescence/luminescence with a plate reader [link part measurement]. Plate
| |
− | readers belong to standard equipment of every lab nowadays, and could deliver easy reproducible results.<br>
| |
− | The second way was to measure the fluorescence by FACS (Fluorescence-Activated Cell Sorting) [link facs]. In
| |
− | contrast to a platerader a FACs device delivers results with high accuracy by measuring every cell by its
| |
− | own(vielleicht erst spaeter FACS genau erklaeren aber nicht im abtract?). On the other side not
| |
− | every laboratory posses a FACs/device. So in the end we would like to offer a two method analyzed database
| |
− | from our crontructs for iGEM teams and research groups, who do not have access to a FACS and show the
| |
− | difference in measurement methods.<br>
| |
− | At the end of the project we were able to create a protocol how to handle Synechococcus elongatus UTEX 2973
| |
− | and make a contribution to the cyano community by establishing essential/fixed standards in measurement.
| |
− | </p>
| |
− | </article>
| |
− | </section>
| |
− | <hr>
| |
− | <section class="section grid">
| |
− | <div class="sub"
| |
− | onclick="popup('rbn1')">
| |
− | <div class="sub-header">
| |
− | <h1>
| |
− | L I G H T<br>
| |
− | M E A S U R E M E N T
| |
− | </h1>
| |
− | <hr>
| |
− | </div>
| |
− | <div class="sub-content">
| |
− | <p>
| |
− | Hier bitte den für diese Stelle zutreffenden Text einfügen, wenn dieser fertig ist.
| |
− | </p>
| |
− | </div>
| |
− | </div>
| |
| <div id="rbn1" | | <div id="rbn1" |
| class="popup"> | | class="popup"> |