title: Reproducibility of Fluorescent Expression from Engineered Biological Constructs in E. coli.
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.
authors.affiliation: Raytheon BBN Technologies, Cambridge, MA, United States of America.
authors.forename: Jacob
authors.initials: J
authors.lastname: Beal
authors.affiliation: iGEM Foundation, Cambridge, MA, United States of America.
authors.forename: Traci
authors.initials: T
authors.lastname: Haddock-Angelli
authors.affiliation: Synthace, London, United Kingdom.
authors.forename: Markus
authors.initials: M
authors.lastname: Gershater
authors.affiliation: iGEM Foundation, Cambridge, MA, United States of America.
authors.forename: Kim
authors.initials: K
authors.lastname: de Mora
authors.affiliation: iGEM Foundation, Cambridge, MA, United States of America.
authors.forename: Meagan
authors.initials: M
authors.lastname: Lizarazo
authors.affiliation: Agilent, Santa Clara, CA, United States of America.
authors.forename: Jim
authors.initials: J
authors.lastname: Hollenhorst
authors.affiliation: iGEM Foundation, Cambridge, MA, United States of America.
authors.forename: Randy
authors.initials: R
authors.lastname: Rettberg
mesh_terms: D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000096:biosynthesis* / Q000235:genetics; D059021:Laboratory Proficiency Testing; D011401:Promoter Regions, Genetic; D015202:Protein Engineering; D015203:Reproducibility of Results; D014158:Transcription, Genetic; D015533:Transcriptional Activation
chemical_list: D049452:Green Fluorescent Proteins