Comparative analysis of three studies measuring fluorescence from engineered bacterial genetic constructs
| Autor: | John P. Marken, Russell Buckley-Taylor, Randy Rettberg, Natalie G. Farny, Marko Storch, Vinoo Selvarajah, Daisuke Kiga, iGEM Interlab Study Contributors, Vishal Sanchania, Meagan Lizarazo, Traci Haddock-Angelli, Christopher T. Workman, Ari Dwijayanti, Abigail Sison, Jacob Beal, Geoff S. Baldwin, Kim de Mora, Markus Gershater |
|---|---|
| Přispěvatelé: | Engineering & Physical Science Research Council (EPSRC), Commission of the European Communities |
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
Luminescence
Standard deviation Spectrum Analysis Techniques 0302 clinical medicine Mathematics 0303 health sciences Multidisciplinary Spectrometers Physics Electromagnetic Radiation Research Assessment Flow Cytometry Fluorescence Reproducibility Multidisciplinary Sciences Spectrophotometry 030220 oncology & carcinogenesis Calibration Physical Sciences Science & Technology - Other Topics Engineering and Technology Medicine Synthetic Biology Cytophotometry Genetic Engineering Biological system Plate reader Research Article Biotechnology Spectrophotometers Engineered genetic General Science & Technology Science Equipment Bioengineering iGEM Interlab Study Contributors Research and Analysis Methods 03 medical and health sciences Genetics Measurement Equipment 030304 developmental biology Science & Technology Bacteria Reproducibility of Results Biology and Life Sciences Sources of error |
| Zdroj: | PLoS ONE, Vol 16, Iss 6, p e0252263 (2021) PLoS ONE PLoS ONE, 16(6) Interlab Study Contributors iGEM, Beal, J, Baldwin, G S, Farny, N G, Gershater, M, Haddock-Angelli, T, Buckley-Taylor, R, Dwijayanti, A, Kiga, D, Lizarazo, M, Marken, J, de Mora, K, Rettberg, R, Sanchania, V, Selvarajah, V, Sison, A, Storch, M & Workman, C T 2021, ' Comparative analysis of three studies measuring fluorescence from engineered bacterial genetic constructs ', PLOS ONE, vol. 16, no. 6, e0252263 . https://doi.org/10.1371/journal.pone.0252263 |
| ISSN: | 1932-6203 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|