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Team:Brasil-SP/Results/interlabstudy
From 2014.igem.org
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<br><span style="font-size: 10-small">Flow cytometry</span> <br><br> | <br><span style="font-size: 10-small">Flow cytometry</span> <br><br> | ||
Revision as of 21:16, 17 October 2014
ProtocolsAs 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
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| Biobrick device 2: BBa_J23101 + BBa_E0240, a new device built by each team Assembly Protocol Biobrick Device 2 and 3
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| Biobrick device 3: BBa_J23115 + BBa_E0240, a new device built by each team. Assembly Protocol Biobrick Device 2 and 3
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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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ResultsFor 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.
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