Pretreatment: Improving endogenous ancient DNA yields using a simple enzymatic predigestion step

Hannes Schroeder*, Peter de Barros Damgaard, Morten E. Allentoft

*Corresponding author for this work
    1 Citation (Scopus)

    Abstract

    Ancient DNA samples generally contain a mixture of both endogenous and exogenous (contaminant) DNA. The authentic endogenous DNA content varies widely between samples and substrates but usually constitutes only a small fraction of the total DNA, while the remainder comprises contamination deriving from bacteria, fungi, and other microorganisms and in some cases also modern human DNA. Recently, several protocols have been developed to improve access to the endogenous DNA fraction by decreasing the exogenous fraction prior to extraction. The most common of these involve pretreatment with single or multiple washes with weak sodium phosphate or sodium hypochlorite (bleach) solutions, as described in Chapter 2. Here, we present an alternative, less aggressive pretreatment protocol that uses a brief predigestion step in an EDTA-based lysis buffer to increase the endogenous fraction prior to extraction.

    Original languageEnglish
    Title of host publicationMethods in Molecular Biology
    Number of pages4
    PublisherHumana Press
    Publication date1 Jan 2019
    Pages21-24
    DOIs
    Publication statusPublished - 1 Jan 2019
    SeriesMethods in Molecular Biology
    Volume1963
    ISSN1064-3745

    Keywords

    • Ancient DNA
    • Contamination
    • EDTA
    • Pretreatment
    • Proteinase K

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