Reproducibility of Fluorescent Expression from Engineered Biological Constructs in E. coli

We present results of the first large-scale interlaboratory study carried out in synthetic biology, as part of the 2014 and 2015 International Genetically Engineered Machine (iGEM) competitions. Participants at 88 institutions around the world measured fluorescence from three engineered constitutive...

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Bibliographic Details
Published in:PloS one 2016-03, Vol.11 (3), p.e0150182-e0150182
Main Authors: Beal, Jacob, Haddock-Angelli, Traci, Gershater, Markus, de Mora, Kim, Lizarazo, Meagan, Hollenhorst, Jim, Rettberg, Randy
Format: Article
Language:English
Subjects:
Analysis
Automation
Bioengineering
Biological effects
Biology and Life Sciences
Calibration
Computer and Information Sciences
Construction engineering
Data processing
E coli
Engineering and Technology
Escherichia coli
Escherichia coli - genetics
Escherichia coli - metabolism
Flow cytometry
Fluorescence
Gene expression
Genetic engineering
Genetically modified organisms
Green Fluorescent Proteins - biosynthesis
Green Fluorescent Proteins - genetics
Laboratories
Laboratory Proficiency Testing
Life Sciences
People and Places
Promoter Regions, Genetic
Protein Engineering
Reproducibility
Reproducibility of Results
Research and Analysis Methods
Synthetic biology
Transcription, Genetic
Transcriptional Activation
Yang, Cindy
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Summary:We present results of the first large-scale interlaboratory study carried out in synthetic biology, as part of the 2014 and 2015 International Genetically Engineered Machine (iGEM) competitions. Participants at 88 institutions around the world measured fluorescence from three engineered constitutive constructs in E. coli. Few participants were able to measure absolute fluorescence, so data was analyzed in terms of ratios. Precision was strongly related to fluorescent strength, ranging from 1.54-fold standard deviation for the ratio between strong promoters to 5.75-fold for the ratio between the strongest and weakest promoter, and while host strain did not affect expression ratios, choice of instrument did. This result shows that high quantitative precision and reproducibility of results is possible, while at the same time indicating areas needing improved laboratory practices.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0150182