Journal article
Quantification of bacterial fluorescence using independent calibrants
PloS one, Vol.13(6), pp.e0199432-e0199432
06/21/2018
DOI: 10.1371/journal.pone.0199432
PMCID: PMC6013168
PMID: 29928012
Abstract
Fluorescent reporters are commonly used to quantify activities or properties of both natural and engineered cells. Fluorescence is still typically reported only in arbitrary or normalized units, however, rather than in units defined using an independent calibrant, which is problematic for scientific reproducibility and even more so when it comes to effective engineering. In this paper, we report an interlaboratory study showing that simple, low-cost unit calibration protocols can remedy this situation, producing comparable units and dramatic improvements in precision over both arbitrary and normalized units. Participants at 92 institutions around the world measured fluorescence from E. coli transformed with three engineered test plasmids, plus positive and negative controls, using simple, low-cost unit calibration protocols designed for use with a plate reader and/or flow cytometer. In addition to providing comparable units, use of an independent calibrant allows quantitative use of positive and negative controls to identify likely instances of protocol failure. The use of independent calibrants thus allows order of magnitude improvements in precision, narrowing the 95% confidence interval of measurements in our study up to 600-fold compared to normalized units.
Details
- Title: Subtitle
- Quantification of bacterial fluorescence using independent calibrants
- Creators
- Jacob Beal - RTXTraci Haddock-Angelli - International Genetically Engineered Machine FoundationGeoff Baldwin - Imperial College LondonMarkus Gershater - SynthaceAri Dwijayanti - Imperial College LondonMarko Storch - Imperial College LondonKim de Mora - International Genetically Engineered Machine FoundationMeagan Lizarazo - International Genetically Engineered Machine FoundationRandy Rettberg - International Genetically Engineered Machine FoundationInternational Genetically Engineered Machine (iGEM) Interlab Study Contributors
- Resource Type
- Journal article
- Publication Details
- PloS one, Vol.13(6), pp.e0199432-e0199432
- DOI
- 10.1371/journal.pone.0199432
- PMID
- 29928012
- PMCID
- PMC6013168
- ISSN
- 1932-6203
- eISSN
- 1932-6203
- Language
- English
- Date published
- 06/21/2018
- Academic Unit
- Electrical and Computer Engineering
- Record Identifier
- 9984627238102771
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