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Assessment of alternatives to environmental toxic formalin for DNA conservation in biological specimens

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Abstract

One essential step of museum and clinical specimen preservation is immersion in a fixative fluid to prevent degradation. Formalin is the most largely used fixative, but its benefit is balanced with its toxic and carcinogenic status. Moreover, because formalin-fixation impairs nucleic acids recovery and quality, current museum wet collections and formalin-fixed, paraffin-embedded clinical samples do not represent optimal tanks of molecular information. Our study has been developed to compare formalin to two alternative fixatives (RCL2® and ethanol) in a context of molecular exploitation. Based on a unique protocol, we created mammalian fixed collections, simulated the impact of time on preservation using an artificial ageing treatment and followed the evolution of specimens’ DNA quality. DNA extraction yield, purity, visual integrity and qualitative and quantitative ability to amplify the Cox1 gene were assessed. Our results show that both RCL2 and ethanol exhibit better performances than formalin. They do not impair DNA extraction yield, and more importantly, DNA alteration is delayed over the preservation step. The use of RCL2 or ethanol as fixative in biological collections may insure a better exploitation of the genetic resources they propose.

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Acknowledgements

This work was supported by PNRCC grant no. 0012994 (Programme national de recherche sur la connaissance et la conservation des matériaux du patrimoine culturel, Ministère de la Culture).

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Authors and Affiliations

  1. INSERM-UMR1037, Université de Toulouse, Centre de Recherches en Cancérologie de Toulouse (CRCT), Equipe Labellisée Ligue Contre le Cancer, Laboratoire d’Excellence Toulouse Cancer (LabEx TOUCAN), 31037, Toulouse, France

    Emeline Sarot & Stéphane Pyronnet

  2. Muséum d’Histoire Naturelle de Toulouse, 35 allées Jules Guesde, 31000, Toulouse, France

    Marie-Françoise Carillo-Baraglioli & Francis Duranthon

  3. Patrimoine Locaux et Gouvernance, UMR208, Muséum national d’Histoire naturelle, 36 rue Geoffroy Saint Hilaire, 75005, Paris, France

    Amandine Péquignot

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  1. Emeline Sarot
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  2. Marie-Françoise Carillo-Baraglioli
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  3. Francis Duranthon
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  4. Amandine Péquignot
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  5. Stéphane Pyronnet
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Correspondence to Stéphane Pyronnet.

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Responsible editor: Philippe Garrigues

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Sarot, E., Carillo-Baraglioli, MF., Duranthon, F. et al. Assessment of alternatives to environmental toxic formalin for DNA conservation in biological specimens. Environ Sci Pollut Res 24, 16985–16993 (2017). https://doi.org/10.1007/s11356-017-9349-y

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