Difference between revisions of "Team:DTU-Denmark/Results-amilCP"

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<div class="headlinecontainer" style="font-size:5vw;"><h1>amilCP Expression</h1></div>
 
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<p>Here you can describe the results of your project and your future plans. </p>
 
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<h3>What should this page contain?</h3>
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<li> Clearly and objectively describe the results of your work.</li>
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<li> Future plans for the project. </li>
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<li> Considerations for replicating the experiments. </li>
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<h3>Describe what your results mean </h3>
 
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<li> Interpretation of the results obtained during your project. Don't just show a plot/figure/graph/other, tell us what you think the data means. This is an important part of your project that the judges will look for. </li>
 
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<li> Show data, but remember all measurement and characterization data must be on part pages in the Registry. </li>
 
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<li> Consider including an analysis summary section to discuss what your results mean. Judges like to read what you think your data means, beyond all the data you have acquired during your project. </li>
 
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<h2 class="media-heading" style="text-align: left;margin-bottom: 35px; color:#50C8E8;">Assembly</h2>
  
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<p>You can also include a list of bullet points (and links) of the successes and failures you have had over your summer. It is a quick reference page for the judges to see what you achieved during your summer.</p>
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AmilCP was acquired from the distribution kit and assembled into pSB1C3 with different fungal/eukaryotic promoters – the two <i>Aspergillus nidulans</i> derived promoters <i>pTrpC</i> and <i>pGpdA</i> as well as the cauliflower mosaic virus 35S promoter - using 3A assembly. Afterwards the terminator from the cauliflower virus was also added via 3A assembly. Correct transformants were identified by colony PCR and Sanger sequencing using the verification primers, VF2 and VR (See figure 1).
  
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<li>A list of linked bullet points of the successful results during your project</li>
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Figure 1: Lav et billede af den relevante gel og inkluder et udklip af sekvensen.
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<li>A list of linked bullet points of the unsuccessful results during your project. This is about being scientifically honest. If you worked on an area for a long time with no success, tell us so we know where you put your effort.</li>
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<h2 class="media-heading"  style="text-align: right;margin-bottom: 35px; color:#F8A05B;">Transformation</h2>
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After confirmed assembly the <i>amilCP</i> expression cassettes were then transformed into <i>Aspergillus oryzae</i> RIB40. Due to the lack of appropriate selectable conditions, none were used. Nonetheless some strains exhibited a darkened spore coloration indicating a possible transformation, see figure 2.
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Figure 2: Tag et billed til sammenligning fra de lineariserede pCaMV
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Genomic DNA was purified from the strains identified as possible transformants. Integration of the <i>amilCP</i> sequence was then confirmed by PCR using VF2 and VR, as well as sequence specific primers, see figure 3.
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Figure 3: Tag gel-billeder med
  
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<h3>Inspiration</h3>
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<p>See how other teams presented their results.</p>
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<li><a href="https://2014.igem.org/Team:TU_Darmstadt/Results/Pathway">2014 TU Darmstadt </a></li>
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<li><a href="https://2014.igem.org/Team:Imperial/Results">2014 Imperial </a></li>
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<li><a href="https://2014.igem.org/Team:Paris_Bettencourt/Results">2014 Paris Bettencourt </a></li>
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<p style="color:#000; font-size:14px;">(1) </p>
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Revision as of 15:39, 16 October 2018

amilCP Expression

Assembly

AmilCP was acquired from the distribution kit and assembled into pSB1C3 with different fungal/eukaryotic promoters – the two Aspergillus nidulans derived promoters pTrpC and pGpdA as well as the cauliflower mosaic virus 35S promoter - using 3A assembly. Afterwards the terminator from the cauliflower virus was also added via 3A assembly. Correct transformants were identified by colony PCR and Sanger sequencing using the verification primers, VF2 and VR (See figure 1).

Figure 1: Lav et billede af den relevante gel og inkluder et udklip af sekvensen.

Transformation

After confirmed assembly the amilCP expression cassettes were then transformed into Aspergillus oryzae RIB40. Due to the lack of appropriate selectable conditions, none were used. Nonetheless some strains exhibited a darkened spore coloration indicating a possible transformation, see figure 2.

Figure 2: Tag et billed til sammenligning fra de lineariserede pCaMV

Genomic DNA was purified from the strains identified as possible transformants. Integration of the amilCP sequence was then confirmed by PCR using VF2 and VR, as well as sequence specific primers, see figure 3.

Figure 3: Tag gel-billeder med

(1)