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

Jacob Beal; Traci Haddock-Angelli; Markus Gershater; Kim de Mora; Meagan Lizarazo; Jim Hollenhorst; Randy Rettberg; IGEM Interlab Study Contributors

doi:10.1371/journal.pone.0150182

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

Journal article

Open access

Peer reviewed

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

Jacob Beal, Traci Haddock-Angelli, Markus Gershater, Kim de Mora, Meagan Lizarazo, Jim Hollenhorst, Randy Rettberg and IGEM Interlab Study Contributors

PloS one, Vol.11(3), pp.e0150182-e0150182

03/03/2016

DOI: 10.1371/journal.pone.0150182

PMCID: PMC4777433

PMID: 26937966

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Abstract

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.

Life Sciences

Details

Title: Subtitle: Reproducibility of Fluorescent Expression from Engineered Biological Constructs in E. coli
Creators: Jacob Beal - RTX
Traci Haddock-Angelli - International Genetically Engineered Machine Foundation
Markus Gershater - Synthace
Kim de Mora - International Genetically Engineered Machine Foundation
Meagan Lizarazo - International Genetically Engineered Machine Foundation
Jim Hollenhorst - Agilent Technologies
Randy Rettberg - International Genetically Engineered Machine Foundation
IGEM Interlab Study Contributors
Resource Type: Journal article
Publication Details: PloS one, Vol.11(3), pp.e0150182-e0150182
Publisher: Public Library of Science
DOI: 10.1371/journal.pone.0150182
PMID: 26937966
PMCID: PMC4777433
ISSN: 1932-6203
eISSN: 1932-6203
Language: English
Date published: 03/03/2016
Academic Unit: Electrical and Computer Engineering
Record Identifier: 9984627222902771

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