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Development of Sensitive Methods for the Detection of Minimum Concentrations of DNA on Martian Soil Simulants

Several methods used for the quantification of DNA are based on UV absorbance or the fluorescence of complexes with intercalator dyes. Most of these intercalators are used in gels to visualize DNA and its structural integrity. Due to many extraterrestrial samples, such as meteorites or comets, which...

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Autores principales: Li, Yongda, Rochfort, Keith D., Collins, David, Grintzalis, Konstantinos
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10608431/
https://www.ncbi.nlm.nih.gov/pubmed/37895382
http://dx.doi.org/10.3390/life13101999
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author Li, Yongda
Rochfort, Keith D.
Collins, David
Grintzalis, Konstantinos
author_facet Li, Yongda
Rochfort, Keith D.
Collins, David
Grintzalis, Konstantinos
author_sort Li, Yongda
collection PubMed
description Several methods used for the quantification of DNA are based on UV absorbance or the fluorescence of complexes with intercalator dyes. Most of these intercalators are used in gels to visualize DNA and its structural integrity. Due to many extraterrestrial samples, such as meteorites or comets, which are likely to contain very small amounts of biological material, and because the ability to detect this material is crucial for understanding the origin and evolution of life in the universe, the development of assays that can detect DNA at low limits and withstand the rigors of space exploration is a pressing need in the field of astrobiology. In this study, we present a comparison of optimized protocols used for the fast and accurate quantification of DNA using common intercalator dyes. The sensitivity of assays exceeded that generated by any commercial kit and allowed for the accurate quantification of minimum concentrations of DNA. The methods were successful when applied to the detection and measurement of DNA spiked on soil samples. Furthermore, the impact of UV radiation as a harsh condition on the surface of Mars was assessed by DNA degradation and this was also confirmed by gel electrophoresis. Overall, the methods described provide economical, simple-step, and efficient approaches for the detection of DNA and can be used in future planetary exploration missions as tests used for the extraction of nucleic acid biosignatures.
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spelling pubmed-106084312023-10-28 Development of Sensitive Methods for the Detection of Minimum Concentrations of DNA on Martian Soil Simulants Li, Yongda Rochfort, Keith D. Collins, David Grintzalis, Konstantinos Life (Basel) Article Several methods used for the quantification of DNA are based on UV absorbance or the fluorescence of complexes with intercalator dyes. Most of these intercalators are used in gels to visualize DNA and its structural integrity. Due to many extraterrestrial samples, such as meteorites or comets, which are likely to contain very small amounts of biological material, and because the ability to detect this material is crucial for understanding the origin and evolution of life in the universe, the development of assays that can detect DNA at low limits and withstand the rigors of space exploration is a pressing need in the field of astrobiology. In this study, we present a comparison of optimized protocols used for the fast and accurate quantification of DNA using common intercalator dyes. The sensitivity of assays exceeded that generated by any commercial kit and allowed for the accurate quantification of minimum concentrations of DNA. The methods were successful when applied to the detection and measurement of DNA spiked on soil samples. Furthermore, the impact of UV radiation as a harsh condition on the surface of Mars was assessed by DNA degradation and this was also confirmed by gel electrophoresis. Overall, the methods described provide economical, simple-step, and efficient approaches for the detection of DNA and can be used in future planetary exploration missions as tests used for the extraction of nucleic acid biosignatures. MDPI 2023-09-30 /pmc/articles/PMC10608431/ /pubmed/37895382 http://dx.doi.org/10.3390/life13101999 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Li, Yongda
Rochfort, Keith D.
Collins, David
Grintzalis, Konstantinos
Development of Sensitive Methods for the Detection of Minimum Concentrations of DNA on Martian Soil Simulants
title Development of Sensitive Methods for the Detection of Minimum Concentrations of DNA on Martian Soil Simulants
title_full Development of Sensitive Methods for the Detection of Minimum Concentrations of DNA on Martian Soil Simulants
title_fullStr Development of Sensitive Methods for the Detection of Minimum Concentrations of DNA on Martian Soil Simulants
title_full_unstemmed Development of Sensitive Methods for the Detection of Minimum Concentrations of DNA on Martian Soil Simulants
title_short Development of Sensitive Methods for the Detection of Minimum Concentrations of DNA on Martian Soil Simulants
title_sort development of sensitive methods for the detection of minimum concentrations of dna on martian soil simulants
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10608431/
https://www.ncbi.nlm.nih.gov/pubmed/37895382
http://dx.doi.org/10.3390/life13101999
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