Comparative analysis of three studies measuring fluorescence from engineered bacterial genetic constructs

Jacob Beal; Geoff Baldwin; Natalie Farny; Markus Gershater; Traci Haddock-Angelli; Russell Buckley-Taylor; Ari Dwijayanti; Daisuke Kiga; Meagan Lizarazo; John Marken; Kim de Mora; Randy Rettberg; Vishal Sanchania; Vinoo Selvarajah; Abigail Sison; Marko Storch; Christopher Workman; Manish Kushwaha; IGEM Interlab Study Contributors

doi:10.1371/journal.pone.0252263

Back

Comparative analysis of three studies measuring fluorescence from engineered bacterial genetic constructs

Journal article

Open access

Peer reviewed

Comparative analysis of three studies measuring fluorescence from engineered bacterial genetic constructs

Jacob Beal, Geoff Baldwin, Natalie Farny, Markus Gershater, Traci Haddock-Angelli, Russell Buckley-Taylor, Ari Dwijayanti, Daisuke Kiga, Meagan Lizarazo, John Marken, …

PloS one, Vol.16(6), pp.e0252263-e0252263

06/07/2021

DOI: 10.1371/journal.pone.0252263

PMCID: PMC8183995

PMID: 34097703

Files and links (1)

url

https://doi.org/10.1371/journal.pone.0252263View

Published (Version of record) Open Access

Abstract

Reproducibility is a key challenge of synthetic biology, but the foundation of reproducibility is only as solid as the reference materials it is built upon. Here we focus on the reproducibility of fluorescence measurements from bacteria transformed with engineered genetic constructs. This comparative analysis comprises three large interlaboratory studies using flow cytometry and plate readers, identical genetic constructs, and compatible unit calibration protocols. Across all three studies, we find similarly high precision in the calibrants used for plate readers. We also find that fluorescence measurements agree closely across the flow cytometry results and two years of plate reader results, with an average standard deviation of 1.52-fold, while the third year of plate reader results are consistently shifted by more than an order of magnitude, with an average shift of 28.9-fold. Analyzing possible sources of error indicates this shift is due to incorrect preparation of the fluorescein calibrant. These findings suggest that measuring fluorescence from engineered constructs is highly reproducible, but also that there is a critical need for access to quality controlled fluorescent calibrants for plate readers.

Engineering Sciences

Life Sciences

Details

Title: Subtitle: Comparative analysis of three studies measuring fluorescence from engineered bacterial genetic constructs
Creators: Jacob Beal - RTX (United States)
Geoff Baldwin - Imperial College London
Natalie Farny - Worcester Polytechnic Institute
Markus Gershater - Synthace (United Kingdom)
Traci Haddock-Angelli - International Genetically Engineered Machine Foundation
Russell Buckley-Taylor - Imperial College London
Ari Dwijayanti - Imperial College London
Daisuke Kiga - Waseda University
Meagan Lizarazo - International Genetically Engineered Machine Foundation
John Marken - California Institute of Technology
Kim de Mora - International Genetically Engineered Machine Foundation
Randy Rettberg - International Genetically Engineered Machine Foundation
Vishal Sanchania - Synthace (United Kingdom)
Vinoo Selvarajah - International Genetically Engineered Machine Foundation
Abigail Sison - International Genetically Engineered Machine Foundation
Marko Storch - Imperial College London
Christopher Workman - Technical University of Denmark
Manish Kushwaha - Microbiologie de l’alimentation au service de la santé
IGEM Interlab Study Contributors
Resource Type: Journal article
Publication Details: PloS one, Vol.16(6), pp.e0252263-e0252263
DOI: 10.1371/journal.pone.0252263
PMID: 34097703
PMCID: PMC8183995
NLM abbreviation: PLoS One
ISSN: 1932-6203
eISSN: 1932-6203
Publisher: Public Library of Science
Language: English
Date published: 06/07/2021
Academic Unit: Electrical and Computer Engineering
Record Identifier: 9984627321802771

Metrics

3 Record Views

10 Times Cited - Web of Science