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DNA Stability in Biodosimetry, Pharmacy and DNA Based Data-Storage: Optimal Storage and Handling Conditions

  • DNA long-term stability and integrity is of importance for applications in DNA based bio-dosimetry, data-storage, pharmaceutical quality-control, donor insemination and DNA based functional nanomaterials. Standard protocols for these applications involve repeated freeze-thaw cycles of the DNA, which can cause detrimental damage to the nucleobases, as well as the sugar-phosphate backbone and therefore the whole molecule. Throughout the literature three hypotheses can be found about the underlying mechanisms occurring during freeze-thaw cycles. It is hypothesized that DNA single-strand breaks during freezing can be induced by mechanical stress leading to shearing of the DNA molecule, by acidic pH causing damage through depurination and beta elimination or by the presence of metal ions catalyzing oxidative damage via reactive oxygen species (ROS). Here we test these hypotheses under well defined conditions with plasmid DNA pUC19 in high-purity buffer (1xPBS) at physiological salt andDNA long-term stability and integrity is of importance for applications in DNA based bio-dosimetry, data-storage, pharmaceutical quality-control, donor insemination and DNA based functional nanomaterials. Standard protocols for these applications involve repeated freeze-thaw cycles of the DNA, which can cause detrimental damage to the nucleobases, as well as the sugar-phosphate backbone and therefore the whole molecule. Throughout the literature three hypotheses can be found about the underlying mechanisms occurring during freeze-thaw cycles. It is hypothesized that DNA single-strand breaks during freezing can be induced by mechanical stress leading to shearing of the DNA molecule, by acidic pH causing damage through depurination and beta elimination or by the presence of metal ions catalyzing oxidative damage via reactive oxygen species (ROS). Here we test these hypotheses under well defined conditions with plasmid DNA pUC19 in high-purity buffer (1xPBS) at physiological salt and pH 7.4 conditions, under pH 6 and in the presence of metal ions in combination with the radical scavengers DMSO and Ectoine. The results show for the 2686 bp long plasmid DNA, that neither mechanical stress, nor pH 6 lead to degradation during repeated freeze-thaw cycles. In contrast, the presence of metal ions (Fe2+) leads to degradation of DNA via the production of radical species.zeige mehrzeige weniger

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Metadaten
Autor*innen:Leo CordsmeierORCiD, Marc Benjamin HahnORCiD
Dokumenttyp:Zeitschriftenartikel
Veröffentlichungsform:Verlagsliteratur
Sprache:Englisch
Titel des übergeordneten Werkes (Englisch):ChemBioChem
Jahr der Erstveröffentlichung:2022
Organisationseinheit der BAM:6 Materialchemie
6 Materialchemie / 6.6 Physik und chemische Analytik der Polymere
Veröffentlichende Institution:Bundesanstalt für Materialforschung und -prüfung (BAM)
Herausgeber (Institution):Chemistry Europe European Chemical Societies Publishing
Verlag:Wiley-VCH GmbH
Aufsatznummer:e202200391
Erste Seite:1
Letzte Seite:9
DDC-Klassifikation:Naturwissenschaften und Mathematik / Chemie / Analytische Chemie
Technik, Medizin, angewandte Wissenschaften / Ingenieurwissenschaften / Sanitär- und Kommunaltechnik; Umwelttechnik
Freie Schlagwörter:Biologisches Dosimeter; DNA Dosimeter; Ectoin
AGE; Base damage; Base loss; Beta elimination; Biodosimetry; Buffer; DMSO; DNA; DNA based data storage; DNA data storage; DNA degradation; DNA long term storage; DNA reference material; DNA stability; DNA vortexing; Dosimetry; Ectoine; Fenton Reaction; Freezing; Gel electrophoresis; H2O2; Mechanical stress; Metal ions; Nucleobase; OH radical; Pharmacy; Plasmid DNA; Quality control; ROS; Radical scavenger; Reference material; SSB; SYBR Gold; Strand break; THP(B); Thawing; lN2; pH; pUC19
Themenfelder/Aktivitätsfelder der BAM:Chemie und Prozesstechnik
Umwelt
Umwelt / Umwelt-Material-Interaktionen
DOI:10.1002/cbic.202200391
URN:urn:nbn:de:kobv:b43-557148
Verfügbarkeit des Dokuments:Datei für die Öffentlichkeit verfügbar ("Open Access")
Lizenz (Deutsch):License LogoCreative Commons - CC BY - Namensnennung 4.0 International
Datum der Freischaltung:15.09.2022
Referierte Publikation:Ja
Datum der Eintragung als referierte Publikation:15.09.2022
Schriftenreihen ohne Nummerierung:Wissenschaftliche Artikel der BAM
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