Methods to maximise recovery of environmental DNA from water samples

    Research output: Contribution to journalArticle

    28 Citations (Scopus)
    6 Downloads (Pure)

    Abstract

    The environmental DNA (eDNA) method is a detection technique that is rapidly gaining credibility as a sensitive tool useful in the surveillance and monitoring of invasive and threatened species. Because eDNA analysis often deals with small quantities of short and degraded DNA fragments, methods that maximize eDNA recovery are required to increase detectability. In this study, we performed experiments at different stages of the eDNA analysis to show which combinations of methods give the best recovery rate for eDNA. Using Oriental weatherloach (Misgurnus anguillicaudatus) as a study species, we show that various combinations of DNA capture, preservation and extraction methods can significantly affect DNA yield. Filtration using cellulose nitrate filter paper preserved in ethanol or stored in a -20°C freezer and extracted with the Qiagen DNeasy kit outperformed other combinations in terms of cost and efficiency of DNA recovery. Our results support the recommendation to filter water samples within 24hours but if this is not possible, our results suggest that refrigeration may be a better option than freezing for short-term storage (i.e., 3-5 days). This information is useful in designing eDNA detection of low-density invasive or threatened species, where small variations in DNA recovery can signify the difference between detection success or failure.

    Original languageEnglish
    Pages (from-to)1-22
    Number of pages22
    JournalPLoS One
    Volume12
    Issue number6
    DOIs
    Publication statusPublished - 1 Jun 2017

    Fingerprint

    Recovery
    Water
    DNA
    water
    sampling
    Misgurnus anguillicaudatus
    methodology
    Introduced Species
    Endangered Species
    threatened species
    invasive species
    water filters
    Cypriniformes
    Refrigeration
    Collodion
    filters
    freezers
    monitoring
    refrigeration
    Freezing

    Cite this

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    title = "Methods to maximise recovery of environmental DNA from water samples",
    abstract = "The environmental DNA (eDNA) method is a detection technique that is rapidly gaining credibility as a sensitive tool useful in the surveillance and monitoring of invasive and threatened species. Because eDNA analysis often deals with small quantities of short and degraded DNA fragments, methods that maximize eDNA recovery are required to increase detectability. In this study, we performed experiments at different stages of the eDNA analysis to show which combinations of methods give the best recovery rate for eDNA. Using Oriental weatherloach (Misgurnus anguillicaudatus) as a study species, we show that various combinations of DNA capture, preservation and extraction methods can significantly affect DNA yield. Filtration using cellulose nitrate filter paper preserved in ethanol or stored in a -20°C freezer and extracted with the Qiagen DNeasy kit outperformed other combinations in terms of cost and efficiency of DNA recovery. Our results support the recommendation to filter water samples within 24hours but if this is not possible, our results suggest that refrigeration may be a better option than freezing for short-term storage (i.e., 3-5 days). This information is useful in designing eDNA detection of low-density invasive or threatened species, where small variations in DNA recovery can signify the difference between detection success or failure.",
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    Methods to maximise recovery of environmental DNA from water samples. / Hinlo, Rheyda; Gleeson, Dianne; Lintermans, Mark; Furlan, Elise.

    In: PLoS One, Vol. 12, No. 6, 01.06.2017, p. 1-22.

    Research output: Contribution to journalArticle

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