Difference between revisions of "Team:Marburg/test"

Line 81: Line 81:
 
         </p>
 
         </p>
 
         <p>
 
         <p>
          <br>
+
           While we wanted to establish Syn. elong. as a new chassis for the iGEM community and scientists we wanted to
          <br>
+
           show the best conditions for cultivation and the best measuring method for our parts in UTEX 2973. Therefore
           While we wanted to establish Syn. elong. as a new chassis for the iGEM community and scientists we
+
           we analyzed a big variety of cultivating conditions in measuring growth curves, tried to find a standard in
          wanted to
+
           light measurement, evaluated different reporters???, established a measurement method and compared it to a
           show the best conditions for cultivation and the best measuring method for our parts in UTEX 2973.
+
           already known FACS measurement method (?).
          Therefore
+
        </p>
           we
+
        <p>
          analyzed a big variety of cultivating conditions in measuring growth curves, tried to find a
+
           At the beginning of our project we faced the first question on how to cultivate UTEX at 1500 μE. [quelle].
          standard in
+
           So we had to measure the light conditions in our incubators and while doing this simple task the first
           light
+
           part of standardization began. We discovered that nearly every paper? is using different methods to measure
          measurement, evaluated different reporters???, established a measurement method and compared it to a
+
           their light conditions and that it is a really complex and important procedure. So we got in contact with
           already
+
           cyano and light measurement experts [link IHP] to confront this problem and standardize it. In the following
          known FACS measurement method (?).
+
           popup we show different ways of measurement, their (dis-)advantages and different results depending on the
          <br>
+
           measuring instrument.<br>
          <br>
+
           Not only the light intensity but also a variety of other cultivating parameters needed to be analyzed.
           At the beginning of our project we faced the first question on how to cultivate UTEX at 1500 μE.
+
           In literature and while talking with different experts (IHP), we recognized that small deviations of these
          [quelle].
+
           parameters had a huge impact on the growth speed of Synechococcus elongatus. While establishing UTEX 2973 as
           So we
+
           a new chassis we evaluated this impact on the growth speed and were able to show combinations of parameters
          had to measure the light conditions in our incubators and while doing this simple task the first
+
           that lead to the fastest growth speed.<br>
           part of
+
           Another aspect was measuring the expression and characterize our part. Different possibilities were discussed
          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
 
           and after testing them we decided on two methods in our project (plate reader and FACs). One
           approach
+
           approach was to measure the fluorescence/luminescence with a plate reader [link part measurement]. Plate
          was to
+
           readers belong to standard equipment of every lab nowadays, and could deliver easy reproducible results.<br>
          measure the fluorescence/luminescence with a plate reader [link part measurement]. Plate readers
+
           The second way was to measure the fluorescence by FACS (Fluorescence-Activated Cell Sorting) [link facs]. In
           belong
+
           contrast to a platerader a FACs device delivers results with high accuracy by measuring every cell by its
          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
 
           own(vielleicht erst spaeter FACS genau erklaeren aber nicht im abtract?). On the other side not
           every
+
           every laboratory posses a FACs/device. So in the end we would like to offer a two method analyzed database
          laboratory
+
          from our crontructs for iGEM teams and research groups, who do not have access to a FACS and show the
          posses a FACs/device. So in the end we would like to offer a two method analyzed database from our
+
          difference in measurement methods.<br>
          crontructs
+
           At the end of the project we were able to create a protocol how to handle Synechococcus elongatus UTEX 2973
          for iGEM teams and research groups, who do not have access to a FACS and show the difference in
+
           and make a contribution to the cyano community by establishing essential/fixed standards in measurement.
          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.
+
          <br>
+
 
         </p>
 
         </p>
 
       </section>
 
       </section>

Revision as of 14:02, 19 October 2019

M E A S U R E M E N T


Amplifying new standards in measurement

Vielleicht noch ein allgemeinem abstract zu Messung (vergleiche andere WIKIS)

Storytelling:

We entered this project as the first Marburg iGEM team working with Synechococcus elongatus UTEX 2973, the fastest phototrophic organism. Missing knowledge in handling and cultivation of UTEX 2973 left us in front of many problems and questions. Especially the usage of different media, light conditions and other cultivating and measurement parameters were one of the biggest problems we discovered in scientific papers. Many of these problems are reasoned in the ongoing optimization and development of methods and instruments. Therefore it is hard to hold on to special methods but still standardization is a huge part in synthetic microbiology and necessary to compare results with other scientists and reproduce their data.

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 (?).

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.
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.
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.
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.
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.


Light measurement

Hier bitte den für diese Stelle zutreffenden Text einfügen, wenn dieser fertig ist.


Reporters

Hier bitte den für diese Stelle zutreffenden Text einfügen, wenn dieser fertig ist.


Fluorescence-Activated Cell Sorting (FACS)

Hier bitte den für diese Stelle zutreffenden Text einfügen, wenn dieser fertig ist.


Part Measurement

For our project it was indispensable to establish a measurement workflow that is not only applicable to UTEX 2973 and other cyanobacteria but also has a high throughput.


Growth Curves

Hier bitte den für diese Stelle zutreffenden Text einfügen, wenn dieser fertig ist.