Team:Brasil-USP/interlabstudy
Interlab Study
2nd International InterLab Measurement Study in synthetic biology
In order to characterize standard biological parts, fluorescence-based assays have been widely used with a variety of fluorophores such as fluorescent proteins (FPs). Green fluorescent protein (GFP) is frequently applied as reporter gene to confirm gene expression and analyse its regulation. The Second International InterLab Measurement Study in synthetic biology proposes to detect three promoters activity by measuring and comparing the levels of GFP expression of three devices expressing GFP and controlled by one of the three promoters (see Figure 1). The three promoters - BBa_J23101, BBa_J23106, BBa_J23117 - are from the Anderson library (http://parts.igem.org/Promoters/Catalog/Anderson), a constitutive promoter family with different strengths.1
Figure 1: Interlab devices 1, 2 and 3, in order from top to down.
Introduction and Motivation
In the eleventh iGEM edition happens the second Interlab Study. This study is based on the characterisation of standard biological parts and, as standard parts, it is fundamental to observe reproducibility and repeatability on their behaviour. Since all the teams have the same devices, we should confirm similar data values qualitative and hopefully quantitative for absolute units. Additionally, it will be possible to analyse the results to enlighten the differences between protocols. In this sense, Interlab study hopes to define well-characterised standard parts. We used a plate reader and a flow cytometer to measure fluorescence with biological and technical replicates, fulfilling InterLab study requirements and extra credit opportunity. Results showed mimimi
We understand the importance of this study and how it helps to define standards BioBricks and analyze the difference between labs and protocols.
Results
In the following subsections, we present our fluorescence measurements over time, a cytometry study showing a more microscopy view of how our cells behaved and our calculations of promoter strength using Relative Promoter Units (RPU). Notice that all of our data is publicly available here, and all software developed to calculate the results presented below are available here.
Fluorescence results - plate reader
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Figure N (click for larger view): Fluorescence readings after 11h of experiment, not normalized by OD yet (see data below).
Cytometry
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Relative Promoter Units
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Measuring Promoter Strength using a Camera + Gimp
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Discussions
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Protocols
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Assemble protocol
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Sample preparation protocols
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Plate reader
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Flow cytometer
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Data acquisition protocol
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Plate reader
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Flow cytometer