Team:Brasil-SP/Results/interlabstudy

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<p><h2>Measurement Interlab Study</h2></p>
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<h1>Measurement Interlab Study</h1>
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<h2 align="left"><b>Protocols</b>
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<p align="justify">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;As part of the first international interlab measurement study in synthetic biology, all Measurement Track teams were required to obtain fluorescence data for three specific genetic devices. They are
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<a href="#characterizationprotocols">Protocols</a>
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<p align="center"> Biobrick device 1: BBa_I20260, an existing device </p>
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<a href="https://static.igem.org/mediawiki/2014/c/c1/Assembly_Biobrick_Device_1_Measurement_Brasilsp.pdf">Assembly Protocol Biobrick Device 1
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<a href="#results">Results</a>
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<p align="center"> Biobrick device 2: BBa_J23101 + BBa_E0240, a new device built by each team</p>
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<a href="https://static.igem.org/mediawiki/2014/d/d3/Assembly_Biobrick_Devices_2_and_3_Measurement_Interlab_Study.pdf"> Assembly Protocol Biobrick Device 2 and 3 </a>
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<p align="justify">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;As part of the first international interlab measurement study in synthetic biology, all Measurement Track teams were required to obtain fluorescence data for three specific genetic devices. They are<p/>
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<p align="center"> Biobrick device 3: BBa_J23115 + BBa_E0240, a new device built by each team.<br><strong>Note:</strong> Compared to biobrick device 1 and 2, biobrick device 3 was constructed with a different promoter (BBa_J23115).
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<a href="https://static.igem.org/mediawiki/2014/d/d3/Assembly_Biobrick_Devices_2_and_3_Measurement_Interlab_Study.pdf"> Assembly Protocol Biobrick Device 2 and 3
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<p align="center"> Biobrick device 1: BBa_I20260, an existing device </p>
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<img src="https://static.igem.org/mediawiki/2014/9/93/Device3_measurement_brasilsp.png" width="350" height="auto">
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<a href="https://static.igem.org/mediawiki/2014/c/c1/Assembly_Biobrick_Device_1_Measurement_Brasilsp.pdf">Assembly Protocol Biobrick Device 1</a>
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<a href="https://static.igem.org/mediawiki/2014/4/40/Interlab_Study_Measurement_Flow_Cytometer_Brasil-SP_Team.pdf">Flow Cytometry Protocol-Samples Preparation
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<p align="justify">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; After colect and store data, it was necessary to analyze our results. Data file format standard for flow cytometry is FCS (Flow Cytometry Standard), it contains all information about data acquired, instrument used and sample. In order to extract useful data we converted .fcs file to .csv using GenePattern Flow Cytometry suite.
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<p align="center"> Biobrick device 2: BBa_J23101 + BBa_E0240, a new device built by each team</p>
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<p align="justify">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
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<a href="https://static.igem.org/mediawiki/2014/d/d3/Assembly_Biobrick_Devices_2_and_3_Measurement_Interlab_Study.pdf"> Assembly Protocol Biobrick Device 2 and 3 </a>
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<h2 align="left"><b>Results</b></h2></ul>
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<p align="justify">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; For the proposed specific genetic devices, we have measured fluorescence by <strong>Flow Cytometry</strong> and <strong>Fluorometry</strong>. Device 1 and 2 are equivalent assemblies, however, we did not expect equal fluorescence values since device 1 was an existing device and 2 was built. When it comes to device 3, only the promoter was changed from BBa_J23101 to BBa_J23115, in all likelihood, differing in strength.
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<span style="font-size: 10-small">Flow cytometry</span>  
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<p align="center"> Biobrick device 3: BBa_J23115 + BBa_E0240, a new device built by each team.<br><strong>Note:</strong> Compared to biobrick device 1 and 2, biobrick device 3 was constructed with a different promoter (BBa_J23115).</p>
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<p align="justify">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Results shown that <strong>highest GFP expression levels</strong> were obtained from biobrick device 1 which promotor was <a href="http://parts.igem.org/Part:BBa_J23101">BBa_J23101</a>, member of a constitutive promoters family whose strongest promoter is J23119. Also from this family, the promoter <a href="http://parts.igem.org/Part:BBa_J23115">BBa_J23115</a>, biobrick device 3,  presented a lower GFP expression level, meaning a <strong>weaker</strong> promoter. Biobrick device 1 and 2 are the same construction, neverthless, because device 2 was constructed, fluorescence was lower.
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<a href="https://static.igem.org/mediawiki/2014/d/d3/Assembly_Biobrick_Devices_2_and_3_Measurement_Interlab_Study.pdf"> Assembly Protocol Biobrick Device 2 and 3 </a>
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    <li><a href="https://static.igem.org/mediawiki/2014/4/40/Interlab_Study_Measurement_Flow_Cytometer_Brasil-SP_Team.pdf">Flow Cytometry Protocol-Samples Preparation</a></li>
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<p align="justify">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; After colect and store data, it was necessary to analyze our results. Data file format standard for flow cytometry is FCS (Flow Cytometry Standard), it contains all information about data acquired, instrument used and sample. In order to extract useful data we converted .fcs file to .csv using GenePattern Flow Cytometry suite.</p>  
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<p align="justify">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; For the proposed specific genetic devices, we have measured fluorescence by <strong>Flow Cytometry</strong> and <strong>Fluorometry</strong>. Device 1 and 2 are equivalent assemblies, however, we did not expect equal fluorescence values since device 1 was an existing device and 2 was built. When it comes to device 3, only the promoter was changed from BBa_J23101 to BBa_J23115, in all likelihood, differing in strength.</p>
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<br><span style="font-size: 10-small">Flow cytometry - Mean values</span>
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Latest revision as of 01:26, 18 October 2014

ResultsBRASILSP.png

Measurement Interlab Study


    Protocols

     As part of the first international interlab measurement study in synthetic biology, all Measurement Track teams were required to obtain fluorescence data for three specific genetic devices. They are

Biobrick device 1: BBa_I20260, an existing device


Assembly Protocol Biobrick Device 1

Biobrick device 2: BBa_J23101 + BBa_E0240, a new device built by each team


Assembly Protocol Biobrick Device 2 and 3


Biobrick device 3: BBa_J23115 + BBa_E0240, a new device built by each team.
Note: Compared to biobrick device 1 and 2, biobrick device 3 was constructed with a different promoter (BBa_J23115).


Assembly Protocol Biobrick Device 2 and 3

      After colect and store data, it was necessary to analyze our results. Data file format standard for flow cytometry is FCS (Flow Cytometry Standard), it contains all information about data acquired, instrument used and sample. In order to extract useful data we converted .fcs file to .csv using GenePattern Flow Cytometry suite.

     

    Results

      For the proposed specific genetic devices, we have measured fluorescence by Flow Cytometry and Fluorometry. Device 1 and 2 are equivalent assemblies, however, we did not expect equal fluorescence values since device 1 was an existing device and 2 was built. When it comes to device 3, only the promoter was changed from BBa_J23101 to BBa_J23115, in all likelihood, differing in strength.



Flow cytometry

     Results shown that highest GFP expression levels were obtained from biobrick device 1 which promotor was BBa_J23101, member of a constitutive promoters family whose strongest promoter is J23119. Also from this family, the promoter BBa_J23115, biobrick device 3, presented a lower GFP expression level, meaning a weaker promoter. Biobrick device 1 and 2 are the same construction, neverthless, because device 2 was constructed, fluorescence was lower.

Sponsors

This project is part of iGEM 2014

Wiki Links

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Retrieved from "http://2014.igem.org/Team:Brasil-SP/Results/interlabstudy"