Team:Gothenburg/Parts/Results

From 2014.igem.org

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<h4>Part Submission</h4>
<h4>Part Submission</h4>
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The part for the registry submission was not successfully transformed into E.coli. As for the yeast transformations, several protocols to merge the plasmid backbone and the part were performed. Unfortunately no success in the subsequently plating was recognizable. Presumably the very short length of the part might has been one of the problems. To separate very short fragments from other pollutants is problematic and probably cost us even the bronze medal.
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As our construct 2 shows fluorescence in the daughter cells but not in the mother cells we know that the cyclin activation is functioning and also that the degradation tag (D-tag) also is working: The D-tag is our submission to the parts registry.
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Revision as of 21:11, 16 October 2014

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Results

In this section we present our laboratory victories!
Unfortunately, not all parts could be created and successfully transformed.
All the genetic fragments on their own could be amplified and purified. Problems accured during the transformations into yeast. Several different transformation protocols were used but the success rate was disappointing.

Constructs

In the end only the two counting constructs could be isolated.
We were able to successfully transform the corresponding fragments into yeast cells, were the homologous recombinase activity merged them together. The positive results by growth on the selective medium were confirmed by a colony PCR.


Figure 1. UV-Picture of the colony PCR gel.

Finally, we also observed our transformed yeast cells under a fluorescent microscope to verify the function of the construct.


Figure 2. Fluorescent microscopy image of transformed yeast cells with the first construct.

Figure 2 shows clearly yellow fluorescence, stating not only the successful transformation but also confirms the function of the ribozymes. Without the cleavage caused by them the fluorescence protein would not be folded properly and no fluorescence would be detactable.

Part Submission

As our construct 2 shows fluorescence in the daughter cells but not in the mother cells we know that the cyclin activation is functioning and also that the degradation tag (D-tag) also is working: The D-tag is our submission to the parts registry.