Fluorescent amplified fragment length polymorphism (fAFLP) analysis of listeria monocytogenes

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    Abstract

    Fluorescent amplified fragment length polymorphism (fAFLP) is based on the selective PCR amplification of restriction fragments from a digest of total genomic DNA. Genomic DNA extracted from a purified bacterial isolate is completely digested with two endonucleases generating fragments which are ligated to specific double-stranded adaptors. The ligated fragments are then amplified by PCR using fluorescently labelled primers. Fluorescent amplified fragments are separated by size on an automated sequencer with a size standard. fAFLP is a rapid, highly reproducible technique which can be used to discriminate and subtype Listeria monocytogenes strains.

    Original languageEnglish
    Pages (from-to)95-101
    Number of pages7
    JournalMethods in molecular biology (Clifton, N.J.)
    Volume1157
    DOIs
    Publication statusPublished - 2014

    Bibliographical note

    Publisher Copyright:
    © Springer Science+Business Media New York 2014.

    Keywords

    • AFLP
    • Fragment analysis
    • Listeria monocytogenes
    • Restriction

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