In:
PLOS ONE, Public Library of Science (PLoS), Vol. 16, No. 6 ( 2021-6-7), p. e0252263-
Kurzfassung:
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.
Materialart:
Online-Ressource
ISSN:
1932-6203
DOI:
10.1371/journal.pone.0252263
DOI:
10.1371/journal.pone.0252263.g001
DOI:
10.1371/journal.pone.0252263.g002
DOI:
10.1371/journal.pone.0252263.g003
DOI:
10.1371/journal.pone.0252263.g004
DOI:
10.1371/journal.pone.0252263.g005
DOI:
10.1371/journal.pone.0252263.s001
DOI:
10.1371/journal.pone.0252263.s002
DOI:
10.1371/journal.pone.0252263.s003
DOI:
10.1371/journal.pone.0252263.s004
DOI:
10.1371/journal.pone.0252263.s005
DOI:
10.1371/journal.pone.0252263.s006
DOI:
10.1371/journal.pone.0252263.s007
DOI:
10.1371/journal.pone.0252263.s008
DOI:
10.1371/journal.pone.0252263.s009
DOI:
10.1371/journal.pone.0252263.s010
DOI:
10.1371/journal.pone.0252263.r001
DOI:
10.1371/journal.pone.0252263.r002
DOI:
10.1371/journal.pone.0252263.r003
DOI:
10.1371/journal.pone.0252263.r004
Sprache:
Englisch
Verlag:
Public Library of Science (PLoS)
Publikationsdatum:
2021
ZDB Id:
2267670-3
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