Difference between revisions of "Team:iTesla-SoundBio/Results"

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<h1>Results</h1>
 
<h1>Results</h1>
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<p>We ultimately sent in two different biobricks: the upstream region of genes pcbA4/5 (BBa_K2363001), and the pcbA5 gene itself (BBa_K2363000). Our control was the digested backbone vector without insert added in the ligation reaction. The plate showed several red colonies and one white colony. We expected nothing to show up, but hey stuff happens. Our plate containing our promoter biobrick had significant growth of white colonies, suggesting the ligations and transformations were effective, so we went on to miniprep and sequence four of the white colonies to make sure they weren’t false positives. There were a few red colonies on the plate which suggests that the digest wasn’t 100% effective and the RFP protein was still expressed. Our pcbA5 biobrick didn’t show as much growth, and there were more red colonies than white colonies. Nevertheless, we went on and miniprepped and sequenced two of the white colonies.</p>
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<p>We ultimately sent in two different biobricks: the upstream region of genes pcbA4/5 (BBa_K2363001) and the pcbA5 gene itself (BBa_K2363000). Our control was the digested backbone vector without insert added in the ligation reaction. With 100% efficiency, we would not expect any colonies to grow on the control plate, but the level of background we observed was minimal. The BBa_K2363001 transformation plate had significant number of white colonies, suggesting the ligations and transformations were effective. We chose to miniprep and sequence two of the white colonies to make sure they were not false positives. There were a few red colonies on the plate which was consistent with the background observed in our control plate. Our pcbA5 biobrick didn’t show as many colonies, and there were more red colonies than white colonies. Nevertheless, we went on to miniprep and sequence two of the white colonies. Sequencing revealed that both the promoter and pcbA5 biobricks obtained from our selected colonies perfectly matched our expected constructs. No point mutations were observed. See raw data attached:</p>
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<a href="https://2017.igem.org/File:T--iTesla-SoundBio--PromSequencing.zip"></a>
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<a href="https://2017.igem.org/File:T--iTesla-SoundBio--A5sequencing.zip"></a>
 
<p>
 
<p>
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<img src="https://static.igem.org/mediawiki/2017/3/3c/T--iTesla-SoundBio--results3.png" align="left" width="300px" /></p>
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<img src="https://static.igem.org/mediawiki/2017/3/3c/T--iTesla-SoundBio--results3.png" align="center" width="300px" /></p>
 
<p>Testing, testing, 1 2 ,3</p><br clear="all" />
 
<p>Testing, testing, 1 2 ,3</p><br clear="all" />
  
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<p><img src="https://static.igem.org/mediawiki/2017/b/b7/T--iTesla-SoundBio--results1.png" align="left" width="300px" /></p>
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<p><img src="https://static.igem.org/mediawiki/2017/b/b7/T--iTesla-SoundBio--results1.png" align="center" width="300px" /></p>
 
<p>ABC: Testing, testing, 1 2 ,3</p><br clear="all" />
 
<p>ABC: Testing, testing, 1 2 ,3</p><br clear="all" />
  
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<p><img src="https://static.igem.org/mediawiki/2017/0/0b/T--iTesla-SoundBio--results2.png" align="left" width="300px" /></p>
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<p><img src="https://static.igem.org/mediawiki/2017/0/0b/T--iTesla-SoundBio--results2.png" align="center" width="300px" /></p>
 
<p>123: Testing, testing, 1 2 ,3</p><br clear="all" />
 
<p>123: Testing, testing, 1 2 ,3</p><br clear="all" />
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<br>
 
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<br>
 
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Sequencing revealed that both the promoter and pcbA5 biobricks obtained from our selected colonies perfectly matched our expected constructs. See raw data attached:
 
 
<br>
 
<br>
 
<br>
 
<br>

Revision as of 02:51, 31 October 2017

Results

We ultimately sent in two different biobricks: the upstream region of genes pcbA4/5 (BBa_K2363001) and the pcbA5 gene itself (BBa_K2363000). Our control was the digested backbone vector without insert added in the ligation reaction. With 100% efficiency, we would not expect any colonies to grow on the control plate, but the level of background we observed was minimal. The BBa_K2363001 transformation plate had significant number of white colonies, suggesting the ligations and transformations were effective. We chose to miniprep and sequence two of the white colonies to make sure they were not false positives. There were a few red colonies on the plate which was consistent with the background observed in our control plate. Our pcbA5 biobrick didn’t show as many colonies, and there were more red colonies than white colonies. Nevertheless, we went on to miniprep and sequence two of the white colonies. Sequencing revealed that both the promoter and pcbA5 biobricks obtained from our selected colonies perfectly matched our expected constructs. No point mutations were observed. See raw data attached:


Testing, testing, 1 2 ,3


ABC: Testing, testing, 1 2 ,3


123: Testing, testing, 1 2 ,3