Isolation and Staining Reveal the Presence of Extracellular DNA in Marine Gel Particles
Abstract
:1. Introduction
2. Results
2.1. Microscopy of Purified Natural MGPs in Liquid Suspension
2.2. Visualisation of the Natural MGP Matrix
2.3. Visualisation of eDNA in Natural MGP Matrix
2.4. A Pseudoalteromonas atlantica MGP Model
2.5. Visualisation of the P. atlantica MGP Model Matrix
2.6. Visualisation of eDNA in P. atlantica EPS (MGP Model)
3. Discussion
3.1. Isolation of MGPs in Liquid Suspension
3.2. The Presence of eDNA in Natural MGPs and P. atlantica MGPs
4. Conclusions
5. Materials and Methods
5.1. Seawater Collection
5.2. Isolation of Natural MGPs from Seawater by Filtration
5.3. Isolation of Concentrated Natural MGPs in Suspension
5.4. Bacterial Strains and Growth Conditions
5.5. A Model MGP Using Pseudoalteromonas atlantica
5.6. Compositional Analysis of Natural MGPs and a P. atlantica MGP Model
5.7. MGP Staining with eDNA Probes
5.8. MGP Visualisation by Microscopy
5.9. Bioimaging
5.10. Quantification of Fluorescent Dye Intensity
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Fluorophore | Target | Excitation (nm) | Acquisition (nm) |
---|---|---|---|
YOYO-1 | eDNA | 491 | 501–532 |
Nile Red | Cell membrane | 553 | 567–781 |
Concavalian A (TMR) | Glycoproteins | 552 | 560–643 |
TOTO-3 | eDNA | 642 | 652–784 |
SYTO-9 | DNA | 483 | 490–591 |
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Al-Wahaibi, A.S.M.; Upstill-Goddard, R.C.; Burgess, J.G. Isolation and Staining Reveal the Presence of Extracellular DNA in Marine Gel Particles. Gels 2023, 9, 251. https://doi.org/10.3390/gels9030251
Al-Wahaibi ASM, Upstill-Goddard RC, Burgess JG. Isolation and Staining Reveal the Presence of Extracellular DNA in Marine Gel Particles. Gels. 2023; 9(3):251. https://doi.org/10.3390/gels9030251
Chicago/Turabian StyleAl-Wahaibi, Aisha S. M., Robert C. Upstill-Goddard, and J. Grant Burgess. 2023. "Isolation and Staining Reveal the Presence of Extracellular DNA in Marine Gel Particles" Gels 9, no. 3: 251. https://doi.org/10.3390/gels9030251