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

Corinne Amar

doi:10.1007/978-1-4939-0703-8_8

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

Corinne Amar

Gastrointestinal Bacteria WGS

Research output: Contribution to journal › Article › peer-review

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 language	English
Pages (from-to)	95-101
Number of pages	7
Journal	Methods in molecular biology (Clifton, N.J.)
Volume	1157
DOIs	https://doi.org/10.1007/978-1-4939-0703-8_8
Publication status	Published - 2014

Bibliographical note

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

Keywords

AFLP
Fragment analysis
Listeria monocytogenes
Restriction

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10.1007/978-1-4939-0703-8_8

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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.",

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AB - 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.

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KW - Restriction

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

Abstract

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Keywords

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